PYRIDO[3,2-d]PYRIMIDINE PI3K DELTA INHIBITOR COMPOUNDS AND METHODS OF USE

ABSTRACT

Formula I compounds, including stereoisomers, geometric isomers, tautomers, metabolites and pharmaceutically acceptable salts thereof, are useful for inhibiting the delta isoform of PI3K, and for treating disorders mediated by lipid kinases such as inflammation, immunological disorders, and cancer. Methods of using compounds of Formula I for in vitro, in situ, and in vivo diagnosis, prevention or treatment of such disorders in mammalian cells, or associated pathological conditions, are disclosed.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. Ser. No. 13/030,246 filed on 18Feb. 2011, and claims the benefit under 35 USC §119(e) of U.S.Provisional Application Ser. No. 61/306,751 filed on 22 Feb. 2010, whichare each incorporated by reference in their entirety.

FIELD OF THE INVENTION

The invention relates generally to compounds for treating disordersmediated by lipid kinases such as inflammation, immunological, andcancer, and more specifically to compounds which inhibit PI3 kinaseactivity. The invention also relates to methods of using the compoundsfor in vitro, in situ, and in vivo diagnosis or treatment of mammaliancells, or associated pathological conditions.

BACKGROUND OF THE INVENTION

Phosphatidylinositol (PI), a phospholipid found in cell membranes, playsan important role in intracellular signal transduction. Cell signalingvia 3′-phosphorylated phosphoinositides has been implicated in a varietyof cellular processes, e.g., malignant transformation, growth factorsignaling, inflammation, and immunity (Rameh et al (1999) J. Biol Chem,274:8347-8350). The enzyme responsible for generating thesephosphorylated signaling products, phosphatidylinositol 3-kinase (alsoreferred to as PI 3-kinase or PI3K), was originally identified as anactivity associated with viral oncoproteins and growth factor receptortyrosine kinases that phosphorylate phosphatidylinositol (PI) and itsphosphorylated derivatives at the 3′-hydroxyl of the inositol ring(Panayotou et al (1992) Trends Cell Biol 2:358-60).

Phosphoinositide 3-kinases (PI3K) are lipid kinases that phosphorylatelipids at the 3-hydroxyl residue of the inositol ring ofphosphoinositols (Whitman et al (1988) Nature, 332:664). The3′-phosphorylated phospholipids (PIP3s) generated by PI3-kinases act assecond messengers recruiting kinases with lipid binding domains(including plekstrin homology (PH) regions), such as Akt andphosphoinositide-dependent kinase-1 (PDK1). Binding of Akt to membranePIP3s causes the translocation of Akt to the plasma membrane, bringingAkt into contact with PDK1, which is responsible for activating Akt. Thetumor-suppressor phosphatase, PTEN, dephosphorylates PIP3 and thereforeacts as a negative regulator of Akt activation. The PI3-kinases Akt andPDK1 are important in the regulation of many cellular processesincluding cell cycle regulation, proliferation, survival, apoptosis andmotility and are significant components of the molecular mechanisms ofdiseases such as cancer, diabetes and immune inflammation (Vivanco et al(2002) Nature Rev. Cancer 2:489; Phillips et al (1998) Cancer 83:41).PI3K inhibition is a promising mechanism for targeted therapies forcancer treatment (Maira et al (2009) Biochem. Soc. Trans. 37:265-272).

PI3 kinase is a heterodimer consisting of p85 and p110 subunits (Otsu etal (1991) Cell 65:91-104; Hiles et al (1992) Cell 70:419-29). Fourdistinct Class I PI3Ks have been identified, designated PI3K α(alpha), β(beta), δ (delta), and γ (gamma), each consisting of a distinct 110 kDacatalytic subunit and a regulatory subunit. More specifically, three ofthe catalytic subunits, i.e., p110 alpha, p110 beta and p110 delta, eachinteract with the same regulatory subunit, p85; whereas p110 gammainteracts with a distinct regulatory subunit, p101. The patterns ofexpression of each of these PI3Ks in human cells and tissues are alsodistinct.

The p110 delta isoform has been implicated in biological functionsrelated to immune-inflammatory diseases, including signaling from theB-cell receptor, T cell receptor, FcR signaling of mast cells andmonocyte/macrophage, and osteoclast function/RANKL signaling (Deane Jand Fruman D A (2004) Annu Rev. Immunol. 2004. 22:563-98; Janas et al.,The Journal of Immunology, 2008, 180: 739-746; Marone R et al., Biochim.Biophy. Acta 2007, 1784:159-185). Deletion of the PI3K delta gene orselective introduction of a catalytically inactive mutant of PI3K deltacauses a nearly complete ablation of B cell proliferation and signaling,and impairment of signaling through T cells as well.

SUMMARY OF THE INVENTION

The invention relates to pyrido[3,2-d]pyrimidine Formula I compoundswith PI3 kinase inhibitory activity and selective binding to the p110delta isoform relative to binding to the p110 alpha isoform.

Formula I compounds have the structures:

and stereoisomers, geometric isomers, tautomers, or pharmaceuticallyacceptable salts thereof. The various substituents, including R¹, R²,R³, R⁴, and R⁵, are as defined herein.

Another aspect of the invention provides a pharmaceutical compositioncomprising a Formula I compound and a pharmaceutically acceptablecarrier, glidant, diluent, or excipient.

The invention also relates to methods of using the Formula I compoundsfor in vitro, in situ, and in vivo diagnosis or treatment of mammaliancells, organisms, or associated pathological conditions, such as cancer,systemic and local inflammation, immune-inflammatory diseases such asrheumatoid arthritis, immune suppression, organ transplant rejection,allergies, ulcerative colitis, Crohn's disease, dermatitis, asthma,systemic lupus erythematosus, Sjögren's Syndrome, multiple sclerosis,scleroderma/systemic sclerosis, idiopathic thrombocytopenic purpura(ITP), anti-neutrophil cytoplasmic antibodies (ANCA) vasculitis, chronicobstructive pulmonary disease (COPD), psoriasis, and for general jointprotective effects.

Another aspect of the invention provides a method of treating a diseaseor disorder which method comprises administering a Formula I compound toa patient with a disease or disorder selected from cancer, immunedisorders, cardiovascular disease, viral infection, inflammation,metabolism/endocrine function disorders and neurological disorders, andmediated by the p110 delta, beta, or alpha isoform of PI3 kinase. Themethod may further comprise administering an additional therapeuticagent selected from a chemotherapeutic agent, an anti-inflammatoryagent, an immunomodulatory agent, a neurotropic factor, an agent fortreating cardiovascular disease, an agent for treating liver disease, ananti-viral agent, an agent for treating blood disorders, an agent fortreating diabetes, and an agent for treating immunodeficiency disorders.

Another aspect of the invention provides a kit for treating a conditionmediated by the p110 delta isoform of PI3 kinase, comprising apharmaceutical composition of a Formula I compound; and instructions foruse.

Other aspects of the invention include: (i) method for preventing ortreating conditions, disorders or diseases mediated by the activation ofthe PI3K kinase enzyme, in a subject in need of such treatment, whichmethod comprises administering to said subject an effective amount of acompound of Formula I or a pharmaceutically acceptable salt thereof, infree form or in a pharmaceutically acceptable salt form as apharmaceutical, in any of the methods as indicated herein; (ii) acompound of the Formula I in free form or in pharmaceutically acceptablesalt form for use as a pharmaceutical in any of the methods describedherein, in particular for the use in one or more phosphatidylinositol3-kinase (PI3K) mediated diseases; (iii) the use of a compound ofFormula I in free form or in pharmaceutically acceptable salt form inany of the methods as indicated herein, in particular for the treatmentof one or more phosphatidylinositol 3-kinase mediated diseases; (iv) theuse of a compound of Formula I in free form or in pharmaceuticallyacceptable salt form in any of the methods as indicated herein, inparticular for the manufacture of a medicament for the treatment of oneor more phosphatidylinositol 3-kinase mediated diseases.

DETAILED DESCRIPTION OF EXEMPLARY EMBODIMENTS

Reference will now be made in detail to certain embodiments of theinvention, examples of which are illustrated in the accompanyingstructures and formulas. While the invention will be described inconjunction with the enumerated embodiments, it will be understood thatthey are not intended to limit the invention to those embodiments. Onthe contrary, the invention is intended to cover all alternatives,modifications, and equivalents which may be included within the scope ofthe present invention as defined by the claims. One skilled in the artwill recognize many methods and materials similar or equivalent to thosedescribed herein, which could be used in the practice of the presentinvention. The present invention is in no way limited to the methods andmaterials described. In the event that one or more of the incorporatedliterature, patents, and similar materials differs from or contradictsthis application, including but not limited to defined terms, termusage, described techniques, or the like, this application controls.

DEFINITIONS

The term “alkyl” as used herein refers to a saturated linear orbranched-chain monovalent hydrocarbon radical of one to twelve carbonatoms (C₁-C₁₂), wherein the alkyl radical may be optionally substitutedindependently with one or more substituents described below. In anotherembodiment, an alkyl radical is one to eight carbon atoms (C₁-C₈), orone to six carbon atoms (C₁-C₆). Examples of alkyl groups include, butare not limited to, methyl (Me, —CH₃), ethyl (Et, —CH₂CH₃), 1-propyl(n-Pr, n-propyl, —CH₂CH₂CH₃), 2-propyl (i-Pr, i-propyl, —CH(CH₃)₂),1-butyl (n-Bu, n-butyl, —CH₂CH₂CH₂CH₃), 2-methyl-1-propyl (i-Bu,i-butyl, —CH₂CH(CH₃)₂), 2-butyl (s-Bu, s-butyl, —CH(CH₃)CH₂CH₃),2-methyl-2-propyl (t-Bu, t-butyl, —C(CH₃)₃), 1-pentyl (n-pentyl,—CH₂CH₂CH₂CH₂CH₃), 2-pentyl(-CH(CH₃)CH₂CH₂CH₃), 3-pentyl(-CH(CH₂CH₃)₂),2-methyl-2-butyl(-C(CH₃)₂CH₂CH₃), 3-methyl-2-butyl(-CH(CH₃)CH(CH₃)₂),3-methyl-1-butyl(-CH₂CH₂CH(CH₃)₂), 2-methyl-1-butyl(-CH₂CH(CH₃)CH₂CH₃),1-hexyl(-CH₂CH₂CH₂CH₂CH₂CH₃), 2-hexyl(-CH(CH₃)CH₂CH₂CH₂CH₃),3-hexyl(-CH(CH₂CH₃)(CH₂CH₂CH₃)), 2-methyl-2-pentyl(-C(CH₃)₂CH₂CH₂CH₃),3-methyl-2-pentyl(-CH(CH₃)CH(CH₃)CH₂CH₃),4-methyl-2-pentyl(-CH(CH₃)CH₂CH(CH₃)₂),3-methyl-3-pentyl(-C(CH₃)(CH₂CH₃)₂),2-methyl-3-pentyl(-CH(CH₂CH₃)CH(CH₃)₂),2,3-dimethyl-2-butyl(-C(CH₃)₂CH(CH₃)₂),3,3-dimethyl-2-butyl(-CH(CH₃)C(CH₃)₃, 1-heptyl, 1-octyl, and the like

The term “alkylene” as used herein refers to a saturated linear orbranched-chain divalent hydrocarbon radical of one to twelve carbonatoms (C₁-C₁₂), wherein the alkylene radical may be optionallysubstituted independently with one or more substituents described below.In another embodiment, an alkylene radical is one to eight carbon atoms(C₁-C₈), or one to six carbon atoms (C₁-C₆). Examples of alkylene groupsinclude, but are not limited to, methylene(-CH₂—), ethylene(-CH₂CH₂—),propylene(-CH₂CH₂CH₂—), and the like.

The term “alkenyl” refers to linear or branched-chain monovalenthydrocarbon radical of two to eight carbon atoms (C₂-C₈) with at leastone site of unsaturation, i.e., a carbon-carbon, sp² double bond,wherein the alkenyl radical may be optionally substituted independentlywith one or more substituents described herein, and includes radicalshaving “cis” and “trans” orientations, or alternatively, “E” and “Z”orientations. Examples include, but are not limited to, ethylenyl orvinyl(-CH═CH₂), allyl(-CH₂CH═CH₂), and the like.

The term “alkenylene” refers to linear or branched-chain divalenthydrocarbon radical of two to eight carbon atoms (C₂-C₈) with at leastone site of unsaturation, i.e., a carbon-carbon, sp² double bond,wherein the alkenyl radical may be optionally substituted, independentlywith one or more substituents described herein and includes radicalshaving “cis” and “trans” orientations, or alternatively, “E” and “Z”orientations. Examples include, but are not limited to, ethylenylene orvinylene(-CH═CH—), allyl(-CH₂CH═CH—), and the like.

The term “alkynyl” refers to a linear or branched monovalent hydrocarbonradical of two to eight carbon atoms (C₂-C₈) with at least one site ofunsaturation, i.e., a carbon-carbon, sp triple bond, wherein the alkynylradical may be optionally substituted independently with one or moresubstituents described herein. Examples include, but are not limited to,ethynyl(-C≡CH), propynyl (propargyl, —CH₂C≡CH), and the like.

The term “alkynylene” refers to a linear or branched divalenthydrocarbon radical of two to eight carbon atoms (C₂-C₈) with at leastone site of unsaturation, i.e., a carbon-carbon, sp triple bond, whereinthe alkynyl radical may be optionally substituted independently with oneor more substituents described herein. Examples include, but are notlimited to, ethynylene(-C≡C—), propynylene (propargylene, —CH₂C≡C—), andthe like.

The terms “carbocycle”, “carbocyclyl”, “carbocyclic ring” and“cycloalkyl” refer to a monovalent non-aromatic, saturated or partiallyunsaturated ring having 3 to 12 carbon atoms (C₃-C₁₂) as a monocyclicring or 7 to 12 carbon atoms as a bicyclic ring. Bicyclic carbocycleshaving 7 to 12 atoms can be arranged, for example, as a bicyclo[4,5],[5,5], [5,6] or [6,6] system, and bicyclic carbocycles having 9 or 10ring atoms can be arranged as a bicyclo[5,6] or [6,6] system, or asbridged systems such as bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane andbicyclo[3.2.2]nonane. Examples of monocyclic carbocycles include, butare not limited to, cyclopropyl, cyclobutyl, cyclopentyl,1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl,1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl,cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl,cycloundecyl, cyclododecyl, and the like.

“Aryl” means a monovalent aromatic hydrocarbon radical of 6-20 carbonatoms (C₆-C₂₀) derived by the removal of one hydrogen atom from a singlecarbon atom of a parent aromatic ring system. Some aryl groups arerepresented in the exemplary structures as “Ar”. Aryl includes bicyclicradicals comprising an aromatic ring fused to a saturated, partiallyunsaturated ring, or aromatic carbocyclic ring. Typical aryl groupsinclude, but are not limited to, radicals derived from benzene (phenyl),substituted benzenes, naphthalene, anthracene, biphenyl, indenyl,indanyl, 1,2-dihydronaphthalene, 1,2,3,4-tetrahydronaphthyl, and thelike. Aryl groups are optionally substituted independently with one ormore substituents described herein.

“Arylene” means a divalent aromatic hydrocarbon radical of 6-20 carbonatoms (C₆-C₂₀) derived by the removal of two hydrogen atom from a twocarbon atoms of a parent aromatic ring system. Some arylene groups arerepresented in the exemplary structures as “Ar”. Arylene includesbicyclic radicals comprising an aromatic ring fused to a saturated,partially unsaturated ring, or aromatic carbocyclic ring. Typicalarylene groups include, but are not limited to, radicals derived frombenzene (phenylene), substituted benzenes, naphthalene, anthracene,biphenylene, indenylene, indanylene, 1,2-dihydronaphthalene,1,2,3,4-tetrahydronaphthyl, and the like. Arylene groups are optionallysubstituted independently with one or more substituents describedherein.

The terms “heterocycle,” “heterocyclyl” and “heterocyclic ring” are usedinterchangeably herein and refer to a saturated or a partiallyunsaturated (i.e., having one or more double and/or triple bonds withinthe ring) carbocyclic radical of 3 to about 20 ring atoms in which atleast one ring atom is a heteroatom selected from nitrogen, oxygen,phosphorus and sulfur, the remaining ring atoms being C, where one ormore ring atoms is optionally substituted independently with one or moresubstituents described below. A heterocycle may be a monocycle having 3to 7 ring members (2 to 6 carbon atoms and 1 to 4 heteroatoms selectedfrom N, O, P, and S) or a bicycle having 7 to 10 ring members (4 to 9carbon atoms and 1 to 6 heteroatoms selected from N, O, P, and S), forexample: a bicyclo[4,5], [5,5], [5,6], or [6,6] system. Heterocycles aredescribed in Paquette, Leo A.; “Principles of Modern HeterocyclicChemistry” (W.A. Benjamin, New York, 1968), particularly Chapters 1, 3,4, 6, 7, and 9; “The Chemistry of Heterocyclic Compounds, A series ofMonographs” (John Wiley & Sons, New York, 1950 to present), inparticular Volumes 13, 14, 16, 19, and 28; and J. Am. Chem. Soc. (1960)82:5566. “Heterocyclyl” also includes radicals where heterocycleradicals are fused with a saturated, partially unsaturated ring, oraromatic carbocyclic or heterocyclic ring. Examples of heterocyclicrings include, but are not limited to, pyrrolidinyl, tetrahydrofuranyl,dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl,tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino,thioxanyl, piperazinyl, homopiperazinyl, azetidinyl, oxetanyl,thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl,thiazepinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl,4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl,dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl,dihydroisoquinolinyl, tetrahydroisoquinolinyl,pyrazolidinylimidazolinyl, imidazolidinyl, 3-azabicyco[3.1.0]hexanyl,3-azabicyclo[4.1.0]heptanyl, azabicyclo[2.2.2]hexanyl, 3H-indolylquinolizinyl and N-pyridyl ureas. Spiro moieties are also includedwithin the scope of this definition. Examples of a heterocyclic groupwherein 2 ring carbon atoms are substituted with oxo (═O) moieties arepyrimidinonyl and 1,1-dioxo-thiomorpholinyl. The heterocycle groupsherein are optionally substituted independently with one or moresubstituents described herein.

The term “heteroaryl” refers to a monovalent aromatic radical of 5-, 6-,or 7-membered rings, and includes fused ring systems (at least one ofwhich is aromatic) of 5-20 atoms, containing one or more heteroatomsindependently selected from nitrogen, oxygen, and sulfur. Examples ofheteroaryl groups are pyridinyl (including, for example,2-hydroxypyridinyl), imidazolyl, imidazopyridinyl, pyrimidinyl(including, for example, 4-hydroxypyrimidinyl), pyrazolyl, triazolyl,pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl,oxadiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl,isoquinolinyl, tetrahydroisoquinolinyl, indolyl, benzimidazolyl,benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl,pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl,triazolyl, thiadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl,benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl,quinoxalinyl, naphthyridinyl, and furopyridinyl. Heteroaryl groups areoptionally substituted independently with one or more substituentsdescribed herein.

The heterocycle or heteroaryl groups may be carbon (carbon-linked), ornitrogen (nitrogen-linked) bonded where such is possible. By way ofexample and not limitation, carbon bonded heterocycles or heteroarylsare bonded at position 2, 3, 4, 5, or 6 of a pyridine, position 3, 4, 5,or 6 of a pyridazine, position 2, 4, 5, or 6 of a pyrimidine, position2, 3, 5, or 6 of a pyrazine, position 2, 3, 4, or 5 of a furan,tetrahydrofuran, thiofuran, thiophene, pyrrole or tetrahydropyrrole,position 2, 4, or 5 of an oxazole, imidazole or thiazole, position 3, 4,or 5 of an isoxazole, pyrazole, or isothiazole, position 2 or 3 of anaziridine, position 2, 3, or 4 of an azetidine, position 2, 3, 4, 5, 6,7, or 8 of a quinoline or position 1, 3, 4, 5, 6, 7, or 8 of anisoquinoline.

By way of example and not limitation, nitrogen bonded heterocycles orheteroaryls are bonded at position 1 of an aziridine, azetidine,pyrrole, pyrrolidine, 2-pyrroline, 3-pyrroline, imidazole,imidazolidine, 2-imidazoline, 3-imidazoline, pyrazole, pyrazoline,2-pyrazoline, 3-pyrazoline, piperidine, piperazine, indole, indoline,1H-indazole, benzimidazole, position 2 of a isoindole, or isoindoline,position 4 of a morpholine, and position 9 of a carbazole, orβ-carboline.

The terms “treat” and “treatment” refer to both therapeutic treatmentand prophylactic or preventative measures, wherein the object is toprevent or slow down (lessen) an undesired physiological change ordisorder, such as the development or spread of cancer. For purposes ofthis invention, beneficial or desired clinical results include, but arenot limited to, alleviation of symptoms, diminishment of extent ofdisease, stabilized (i.e., not worsening) state of disease, delay orslowing of disease progression, amelioration or palliation of thedisease state, and remission (whether partial or total), whetherdetectable or undetectable. “Treatment” can also mean prolongingsurvival as compared to expected survival if not receiving treatment.Those in need of treatment include those already with the condition ordisorder as well as those prone to have the condition or disorder orthose in which the condition or disorder is to be prevented.

The phrase “therapeutically effective amount” means an amount of acompound of the present invention that (i) treats or prevents theparticular disease, condition, or disorder, (ii) attenuates,ameliorates, or eliminates one or more symptoms of the particulardisease, condition, or disorder, or (iii) prevents or delays the onsetof one or more symptoms of the particular disease, condition, ordisorder described herein. In the case of cancer, the therapeuticallyeffective amount of the drug may reduce the number of cancer cells;reduce the tumor size; inhibit (i.e., slow to some extent and preferablystop) cancer cell infiltration into peripheral organs; inhibit (i.e.,slow to some extent and preferably stop) tumor metastasis; inhibit, tosome extent, tumor growth; and/or relieve to some extent one or more ofthe symptoms associated with the cancer. To the extent the drug mayprevent growth and/or kill existing cancer cells, it may be cytostaticand/or cytotoxic. For cancer therapy, efficacy can be measured, forexample, by assessing the time to disease progression (TTP) and/ordetermining the response rate (RR).

“Inflammatory disorder” as used herein can refer to any disease,disorder, or syndrome in which an excessive or unregulated inflammatoryresponse leads to excessive inflammatory symptoms, host tissue damage,or loss of tissue function. “Inflammatory disorder” also refers to apathological state mediated by influx of leukocytes and/or neutrophilchemotaxis.

“Inflammation” as used herein refers to a localized, protective responseelicited by injury or destruction of tissues, which serves to destroy,dilute, or wall off (sequester) both the injurious agent and the injuredtissue. Inflammation is notably associated with influx of leukocytesand/or neutrophil chemotaxis. Inflammation can result from infectionwith pathogenic organisms and viruses and from noninfectious means suchas trauma or reperfusion following myocardial infarction or stroke,immune response to foreign antigen, and autoimmune responses.Accordingly, inflammatory disorders amenable to treatment with Formula Icompounds encompass disorders associated with reactions of the specificdefense system as well as with reactions of the nonspecific defensesystem.

“Specific defense system” refers to the component of the immune systemthat reacts to the presence of specific antigens. Examples ofinflammation resulting from a response of the specific defense systeminclude the classical response to foreign antigens, autoimmune diseases,and delayed type hypersensitivity response mediated by T-cells. Chronicinflammatory diseases, the rejection of solid transplanted tissue andorgans, e.g., kidney and bone marrow transplants, and graft versus hostdisease (GVHD), are further examples of inflammatory reactions of thespecific defense system.

The term “nonspecific defense system” as used herein refers toinflammatory disorders that are mediated by leukocytes that areincapable of immunological memory (e.g., granulocytes, and macrophages).Examples of inflammation that result, at least in part, from a reactionof the nonspecific defense system include inflammation associated withconditions such as adult (acute) respiratory distress syndrome (ARDS) ormultiple organ injury syndromes; reperfusion injury; acuteglomerulonephritis; reactive arthritis; dermatoses with acuteinflammatory components; acute purulent meningitis or other centralnervous system inflammatory disorders such as stroke; thermal injury;inflammatory bowel disease; granulocyte transfusion associatedsyndromes; and cytokine-induced toxicity.

“Autoimmune disease” as used herein refers to any group of disorders inwhich tissue injury is associated with humoral or cell-mediatedresponses to the body's own constituents.

“Allergic disease” as used herein refers to any symptoms, tissue damage,or loss of tissue function resulting from allergy. “Arthritic disease”as used herein refers to any disease that is characterized byinflammatory lesions of the joints attributable to a variety ofetiologies. “Dermatitis” as used herein refers to any of a large familyof diseases of the skin that are characterized by inflammation of theskin attributable to a variety of etiologies. “Transplant rejection” asused herein refers to any immune reaction directed against graftedtissue, such as organs or cells (e.g., bone marrow), characterized by aloss of function of the grafted and surrounding tissues, pain, swelling,leukocytosis, and thrombocytopenia. The therapeutic methods of thepresent invention include methods for the treatment of disordersassociated with inflammatory cell activation.

“Inflammatory cell activation” refers to the induction by a stimulus(including, but not limited to, cytokines, antigens or auto-antibodies)of a proliferative cellular response, the production of solublemediators (including but not limited to cytokines, oxygen radicals,enzymes, prostanoids, or vasoactive amines), or cell surface expressionof new or increased numbers of mediators (including, but not limited to,major histocompatability antigens or cell adhesion molecules) ininflammatory cells (including but not limited to monocytes, macrophages,T lymphocytes, B lymphocytes, granulocytes (i.e., polymorphonuclearleukocytes such as neutrophils, basophils, and eosinophils), mast cells,dendritic cells, Langerhans cells, and endothelial cells). It will beappreciated by persons skilled in the art that the activation of one ora combination of these phenotypes in these cells can contribute to theinitiation, perpetuation, or exacerbation of an inflammatory disorder.

The term “NSAID” is an acronym for “non-steroidal anti-inflammatorydrug” and is a therapeutic agent with analgesic, antipyretic (loweringan elevated body temperature and relieving pain without impairingconsciousness) and, in higher doses, with anti-inflammatory effects(reducing inflammation). The term “non-steroidal” is used to distinguishthese drugs from steroids, which (among a broad range of other effects)have a similar eicosanoid-depressing, anti-inflammatory action. Asanalgesics, NSAIDs are unusual in that they are non-narcotic. NSAIDsinclude aspirin, ibuprofen, and naproxen. NSAIDs are usually indicatedfor the treatment of acute or chronic conditions where pain andinflammation are present. NSAIDs are generally indicated for thesymptomatic relief of the following conditions: rheumatoid arthritis,osteoarthritis, inflammatory arthropathies (e.g. ankylosing spondylitis,psoriatic arthritis, Reiter's syndrome, acute gout, dysmenorrhoea,metastatic bone pain, headache and migraine, postoperative pain,mild-to-moderate pain due to inflammation and tissue injury, pyrexia,ileus, and renal colic. Most NSAIDs act as non-selective inhibitors ofthe enzyme cyclooxygenase, inhibiting both the cyclooxygenase-1 (COX-1)and cyclooxygenase-2 (COX-2) isoenzymes. Cyclooxygenase catalyzes theformation of prostaglandins and thromboxane from arachidonic acid(itself derived from the cellular phospholipid bilayer by phospholipaseA₂). Prostaglandins act (among other things) as messenger molecules inthe process of inflammation. COX-2 inhibitors include celecoxib,etoricoxib, lumiracoxib, parecoxib, rofecoxib, rofecoxib, andvaldecoxib.

The terms “cancer” refers to or describe the physiological condition inmammals that is typically characterized by unregulated cell growth. A“tumor” comprises one or more cancerous cells. Examples of cancerinclude, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma,and leukemia or lymphoid malignancies. More particular examples of suchcancers include squamous cell cancer (e.g., epithelial squamous cellcancer), lung cancer including small-cell lung cancer, non-small celllung cancer (“NSCLC”), adenocarcinoma of the lung and squamous carcinomaof the lung, cancer of the peritoneum, hepatocellular cancer, gastric orstomach cancer including gastrointestinal cancer, pancreatic cancer,glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladdercancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectalcancer, endometrial or uterine carcinoma, salivary gland carcinoma,kidney or renal cancer, prostate cancer, vulval cancer, thyroid cancer,hepatic carcinoma, anal carcinoma, penile carcinoma, as well as head andneck cancer.

A “chemotherapeutic agent” is a chemical compound useful in thetreatment of cancer, regardless of mechanism of action. Classes ofchemotherapeutic agents include, but are not limited to: alkylatingagents, antimetabolites, spindle poison plant alkaloids,cytotoxic/antitumor antibiotics, topoisomerase inhibitors, antibodies,photosensitizers, and kinase inhibitors. Chemotherapeutic agents includecompounds used in “targeted therapy” and conventional chemotherapy.Examples of chemotherapeutic agents include: erlotinib (TARCEVA®,Genentech/OSI Pharm.), docetaxel (TAXOTERE®, Sanofi-Aventis), 5-FU(fluorouracil, 5-fluorouracil, CAS No. 51-21-8), gemcitabine (GEMZAR®,Lilly), PD-0325901 (CAS No. 391210-10-9, Pfizer), cisplatin(cis-diamine, dichloroplatinum(II), CAS No. 15663-27-1), carboplatin(CAS No. 41575-94-4), paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology,Princeton, N.J.), trastuzumab (HERCEPTIN®, Genentech), temozolomide(4-methyl-5-oxo-2,3,4,6,8-pentazabicyclo[4.3.0]nona-2,7,9-triene-9-carboxamide, CAS No. 85622-93-1, TEMODAR®, TEMODAL®,Schering Plough), tamoxifen((Z)-2-[4-(1,2-diphenylbut-1-enyl)phenoxy]-N,N-dimethylethanamine,NOLVADEX®, ISTUBAL®, VALODEX®), and doxorubicin (ADRIAMYCIN®), Akti-1/2,HPPD, and rapamycin.

More examples of chemotherapeutic agents include: oxaliplatin(ELOXATIN®, Sanofi), bortezomib (VELCADE®, Millennium Pharm.), sutent(SUNITINIB®, SU11248, Pfizer), letrozole (FEMARA®, Novartis), imatinibmesylate (GLEEVEC®, Novartis), XL-518 (MEK inhibitor, Exelixis, WO2007/044515), ARRY-886 (Mek inhibitor, AZD6244, Array BioPharma, AstraZeneca), SF-1126 (PI3K inhibitor, Semafore Pharmaceuticals), BEZ-235(PI3K inhibitor, Novartis), XL-147 (PI3K inhibitor, Exelixis), PTK787/ZK222584 (Novartis), fulvestrant (FASLODEX®, AstraZeneca), leucovorin(folinic acid), rapamycin (sirolimus, RAPAMUNE®, Wyeth), lapatinib(TYKERB®, GSK572016, Glaxo Smith Kline), lonafarnib (SARASAR™, SCH66336, Schering Plough), sorafenib (NEXAVAR®, BAY43-9006, Bayer Labs),gefitinib (IRESSA®, AstraZeneca), irinotecan (CAMPTOSAR®, CPT-11,Pfizer), tipifarnib (ZARNESTRA™, Johnson & Johnson), ABRAXANE™(Cremophor-free), albumin-engineered nanoparticle formulations ofpaclitaxel (American Pharmaceutical Partners, Schaumberg, II),vandetanib (rINN, ZD6474, ZACTIMA®, AstraZeneca), chloranmbucil, AG1478,AG1571 (SU 5271; Sugen), temsirolimus (TORISEL®, Wyeth), pazopanib(GlaxoSmithKline), canfosfamide (TELCYTA®, Telik), thiotepa andcyclosphosphamide (CYTOXAN®, NEOSAR®); alkyl sulfonates such asbusulfan, improsulfan and piposulfan; aziridines such as benzodopa,carboquone, meturedopa, and uredopa; ethylenimines and methylamelaminesincluding altretamine, triethylenemelamine, triethylenephosphoramide,triethylenethiophosphoramide and trimethylomelamine; acetogenins(especially bullatacin and bullatacinone); a camptothecin (including thesynthetic analog topotecan); bryostatin; callystatin; CC-1065 (includingits adozelesin, carzelesin and bizelesin synthetic analogs);cryptophycins (particularly cryptophycin 1 and cryptophycin 8);dolastatin; duocarmycin (including the synthetic analogs, KW-2189 andCB1-TM1); eleutherobin; pancratistatin; a sarcodictyin; spongistatin;nitrogen mustards such as chlorambucil, chlornaphazine,chlorophosphamide, estramustine, ifosfamide, mechlorethamine,mechlorethamine oxide hydrochloride, melphalan, novembichin,phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureassuch as carmustine, chlorozotocin, fotemustine, lomustine, nimustine,and ranimnustine; antibiotics such as the enediyne antibiotics (e.g.,calicheamicin, calicheamicin gamma1I, calicheamicin omegaI1 (Angew Chem.Intl. Ed. Engl. (1994) 33:183-186); dynemicin, dynemicin A;bisphosphonates, such as clodronate; an esperamicin; as well asneocarzinostatin chromophore and related chromoprotein enediyneantibiotic chromophores), aclacinomysins, actinomycin, authramycin,azaserine, bleomycins, cactinomycin, carabicin, caminomycin,carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin,6-diazo-5-oxo-L-norleucine, morpholino-doxorubicin,cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin anddeoxydoxorubicin), epirubicin, esorubicin, idarubicin, nemorubicin,marcellomycin, mitomycins such as mitomycin C, mycophenolic acid,nogalamycin, olivomycins, peplomycin, porfiromycin, puromycin,quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin,ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexateand 5-fluorouracil (5-FU); folic acid analogs such as denopterin,methotrexate, pteropterin, trimetrexate; purine analogs such asfludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidineanalogs such as ancitabine, azacytidine, 6-azauridine, carmofur,cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine;androgens such as calusterone, dromostanolone propionate, epitiostanol,mepitiostane, testolactone; anti-adrenals such as aminoglutethimide,mitotane, trilostane; folic acid replenisher such as frolinic acid;aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil;amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine;diaziquone; elformithine; elliptinium acetate; an epothilone; etoglucid;gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids suchas maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol;nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone;podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK®polysaccharidecomplex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin;sizofiran; spirogermanium; tenuazonic acid; triaziquone;2,2′,2″-trichlorotriethylamine; trichothecenes (especially T-2 toxin,verracurin A, roridin A and anguidine); urethan; vindesine; dacarbazine;mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine;arabinoside (“Ara-C”); cyclophosphamide; thiotepa; 6-thioguanine;mercaptopurine; methotrexate; platinum analogs such as cisplatin andcarboplatin; vinblastine; etoposide (VP-16); ifosfamide; mitoxantrone;vincristine; vinorelbine (NAVELBINE®); novantrone; teniposide;edatrexate; daunomycin; aminopterin; capecitabine (XELODA®, Roche);ibandronate; CPT-11; topoisomerase inhibitor RFS 2000;difluoromethylornithine (DMFO); retinoids such as retinoic acid; andpharmaceutically acceptable salts, acids and derivatives of any of theabove.

Also included in the definition of “chemotherapeutic agent” are: (i)anti-hormonal agents that act to regulate or inhibit hormone action ontumors such as anti-estrogens and selective estrogen receptor modulators(SERMs), including, for example, tamoxifen (including NOLVADEX®;tamoxifen citrate), raloxifene, droloxifene, 4-hydroxytamoxifen,trioxifene, keoxifene, LY117018, onapristone, and FARESTON® (toremifinecitrate); (ii) aromatase inhibitors that inhibit the enzyme aromatase,which regulates estrogen production in the adrenal glands, such as, forexample, 4(5)-imidazoles, aminoglutethimide, MEGASE® (megestrolacetate), AROMASIN® (exemestane; Pfizer), formestanie, fadrozole,RIVISOR® (vorozole), FEMARA® (letrozole; Novartis), and ARIMIDEX®(anastrozole; AstraZeneca); (iii) anti-androgens such as flutamide,nilutamide, bicalutamide, leuprolide, and goserelin; as well astroxacitabine (a 1,3-dioxolane nucleoside cytosine analog); (iv) proteinkinase inhibitors such as MEK inhibitors (WO 2007/044515); (v) lipidkinase inhibitors; (vi) antisense oligonucleotides, particularly thosewhich inhibit expression of genes in signaling pathways implicated inaberrant cell proliferation, for example, PKC-alpha, Raf and H-Ras, suchas oblimersen (GENASENSE®, Genta Inc.); (vii) ribozymes such as VEGFexpression inhibitors (e.g., ANGIOZYME®) and HER2 expression inhibitors;(viii) vaccines such as gene therapy vaccines, for example, ALLOVECTIN®,LEUVECTIN®, and VAXID®; PROLEUKIN® rIL-2; (ix) topoisomerase 1inhibitors such as LURTOTECAN®; ABARELIX® rmRH; (x) anti-angiogenicagents such as bevacizumab (AVASTIN®, Genentech); (xi) alkylating agentssuch as VNP-40101M or cloretizine, oxaliplatin (U.S. Pat. No. 4,169,846,WO 03/24978m WO 03/04505), glufosfamide, mafosfamide, etopophos (U.S.Pat. No. 5,041,424), prednimustine; treosulfan; busulfan; irofluven(acylfulvene); penclomedine; pyrazoloacridine (PD-115934);06-benzylguanine; decitabine (5-aza-2-deoxycytidine), brostallicin,mitomycin C (MitoExtra), TLK-286 (TELCYTA™), temozolomide, trabectedin(U.S. Pat. No. 5,478,932), AP-5280 (Platinate formulation of Cisplatin),porfiromycin, and clearazide (meclorethamine); (xii) chelating agentsincluding tetrathiomolybdate (WO 01/60814), RP-697, Chimeric T84.66(cT84.66), gadofosveset (VASOVIST™), deferoxamine, and bleomycinoptionally in combination with electroporation; (xiii) anti-cancervaccines including AVICINE™ (Tetrahedron Lett. 26:2269-70 (1974)),oregovomab (OVAREX™), THERATOPE™ (STn-KLH), Melanoma Vaccines, GI-4000series (GI-4014, GI-4015, and GI-4016), directed to mutations in the Rasprotein, GlioVax-1, MelaVax, Advexin™ or INGN-201 (WO 95/12660),Sig/E7/LAMP-1, encoding HPV-16 E7, MAGE-3 Vaccine or M3TK (WO 94/05304),HER-2VAX, ACTIVE which stimulates T-cells specific for tumors, GM-CSFcancer vaccine, ICT-107 (ImmunoCellular Therapeutics), and Listeriamonocytogenes-based vaccines; and pharmaceutically acceptable salts,acids and derivatives of any of the above.

Also included in the definition of “chemotherapeutic agent” aretherapeutic antibodies such as alemtuzumab (Campath), bevacizumab(AVASTIN®, Genentech); cetuximab (ERBITUX®, Imclone); panitumumab(VECTIBIX®, Amgen), rituximab (RITUXAN®, Genentech/Biogen Idec),pertuzumab (OMNITARG™, 2C4, Genentech), trastuzumab (HERCEPTIN®,Genentech), tositumomab (Bexxar, Corixia), and the antibody drugconjugate, gemtuzumab ozogamicin (MYLOTARG®, Wyeth).

Humanized monoclonal antibodies with therapeutic potential aschemotherapeutic agents in combination with the PI3K inhibitors of theinvention include: alemtuzumab, apolizumab, aselizumab, atlizumab,bapineuzumab, bevacizumab, bivatuzumab mertansine, cantuzumabmertansine, cedelizumab, certolizumab pegol, cidfusituzumab, cidtuzumab,daclizumab, eculizumab, efalizumab, epratuzumab, erlizumab, felvizumab,fontolizumab, gemtuzumab ozogamicin, inotuzumab ozogamicin, ipilimumab,labetuzumab, lintuzumab, matuzumab, mepolizumab, motavizumab,motovizumab, natalizumab, nimotuzumab, nolovizumab, numavizumab,ocrelizumab, omalizumab, palivizumab, pascolizumab, pecfusituzumab,pectuzumab, pertuzumab, pexelizumab, ralivizumab, ranibizumab,reslivizumab, reslizumab, resyvizumab, rovelizumab, ruplizumab,sibrotuzumab, siplizumab, sontuzumab, tacatuzumab tetraxetan,tadocizumab, talizumab, tefibazumab, tocilizumab, toralizumab,trastuzumab, tucotuzumab celmoleukin, tucusituzumab, umavizumab,urtoxazumab, and visilizumab.

A “metabolite” is a product produced through metabolism in the body of aspecified compound or salt thereof. Metabolites of a compound may beidentified using routine techniques known in the art and theiractivities determined using tests such as those described herein. Suchproducts may result for example from the oxidation, reduction,hydrolysis, amidation, deamidation, esterification, deesterification,enzymatic cleavage, and the like, of the administered compound.Accordingly, the invention includes metabolites of compounds of theinvention, including compounds produced by a process comprisingcontacting a compound of this invention with a mammal for a period oftime sufficient to yield a metabolic product thereof.

The term “package insert” is used to refer to instructions customarilyincluded in commercial packages of therapeutic products, that containinformation about the indications, usage, dosage, administration,contraindications and/or warnings concerning the use of such therapeuticproducts.

The term “chiral” refers to molecules which have the property ofnon-superimposability of the mirror image partner, while the term“achiral” refers to molecules which are superimposable on their mirrorimage partner.

The term “stereoisomers” refers to compounds which have identicalchemical constitution, but differ with regard to the arrangement of theatoms or groups in space.

“Diastereomer” refers to a stereoisomer with two or more centers ofchirality and whose molecules are not mirror images of one another.Diastereomers have different physical properties, e.g. melting points,boiling points, spectral properties, and reactivities. Mixtures ofdiastereomers may separate under high resolution analytical proceduressuch as electrophoresis and chromatography.

“Enantiomers” refer to two stereoisomers of a compound which arenon-superimposable mirror images of one another.

Stereochemical definitions and conventions used herein generally followS. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984)McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S.,“Stereochemistry of Organic Compounds”, John Wiley & Sons, Inc., NewYork, 1994. The compounds of the invention may contain asymmetric orchiral centers, and therefore exist in different stereoisomeric forms.It is intended that all stereoisomeric forms of the compounds of theinvention, including but not limited to, diastereomers, enantiomers andatropisomers, as well as mixtures thereof such as racemic mixtures, formpart of the present invention. Many organic compounds exist in opticallyactive forms, i.e., they have the ability to rotate the plane ofplane-polarized light. In describing an optically active compound, theprefixes D and L, or R and S, are used to denote the absoluteconfiguration of the molecule about its chiral center(s). The prefixes dand l or (+) and (−) are employed to designate the sign of rotation ofplane-polarized light by the compound, with (−) or 1 meaning that thecompound is levorotatory. A compound prefixed with (+) or d isdextrorotatory. For a given chemical structure, these stereoisomers areidentical except that they are mirror images of one another. A specificstereoisomer may also be referred to as an enantiomer, and a mixture ofsuch isomers is often called an enantiomeric mixture. A 50:50 mixture ofenantiomers is referred to as a racemic mixture or a racemate, which mayoccur where there has been no stereoselection or stereospecificity in achemical reaction or process. The terms “racemic mixture” and “racemate”refer to an equimolar mixture of two enantiomeric species, devoid ofoptical activity.

The term “tautomer” or “tautomeric form” refers to structural isomers ofdifferent energies which are interconvertible via a low energy barrier.For example, proton tautomers (also known as prototropic tautomers)include interconversions via migration of a proton, such as keto-enoland imine-enamine isomerizations. Valence tautomers includeinterconversions by reorganization of some of the bonding electrons.

The phrase “pharmaceutically acceptable salt” as used herein, refers topharmaceutically acceptable organic or inorganic salts of a compound ofthe invention. Exemplary salts include, but are not limited, to sulfate,citrate, acetate, oxalate, chloride, bromide, iodide, nitrate,bisulfate, phosphate, acid phosphate, isonicotinate, lactate,salicylate, acid citrate, tartrate, oleate, tannate, pantothenate,bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate,gluconate, glucuronate, saccharate, formate, benzoate, glutamate,methanesulfonate “mesylate”, ethanesulfonate, benzenesulfonate,p-toluenesulfonate, and pamoate (i.e.,1,1′-methylene-bis(2-hydroxy-3-naphthoate)) salts. A pharmaceuticallyacceptable salt may involve the inclusion of another molecule such as anacetate ion, a succinate ion or other counter ion. The counter ion maybe any organic or inorganic moiety that stabilizes the charge on theparent compound. Furthermore, a pharmaceutically acceptable salt mayhave more than one charged atom in its structure. Instances wheremultiple charged atoms are part of the pharmaceutically acceptable saltcan have multiple counter ions. Hence, a pharmaceutically acceptablesalt can have one or more charged atoms and/or one or more counter ion.

If the compound of the invention is a base, the desired pharmaceuticallyacceptable salt may be prepared by any suitable method available in theart, for example, treatment of the free base with an inorganic acid,such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,methanesulfonic acid, phosphoric acid and the like, or with an organicacid, such as acetic acid, trifluoroacetic acid, maleic acid, succinicacid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalicacid, glycolic acid, salicylic acid, a pyranosidyl acid, such asglucuronic acid or galacturonic acid, an alpha hydroxy acid, such ascitric acid or tartaric acid, an amino acid, such as aspartic acid orglutamic acid, an aromatic acid, such as benzoic acid or cinnamic acid,a sulfonic acid, such as p-toluenesulfonic acid or ethanesulfonic acid,or the like.

If the compound of the invention is an acid, the desiredpharmaceutically acceptable salt may be prepared by any suitable method,for example, treatment of the free acid with an inorganic or organicbase, such as an amine (primary, secondary or tertiary), an alkali metalhydroxide or alkaline earth metal hydroxide, or the like. Illustrativeexamples of suitable salts include, but are not limited to, organicsalts derived from amino acids, such as glycine and arginine, ammonia,primary, secondary, and tertiary amines, and cyclic amines, such aspiperidine, morpholine and piperazine, and inorganic salts derived fromsodium, calcium, potassium, magnesium, manganese, iron, copper, zinc,aluminum and lithium.

The phrase “pharmaceutically acceptable” indicates that the substance orcomposition must be compatible chemically and/or toxicologically, withthe other ingredients comprising a formulation, and/or the mammal beingtreated therewith.

A “solvate” refers to an association or complex of one or more solventmolecules and a compound of the invention. Examples of solvents thatform solvates include, but are not limited to, water, isopropanol,ethanol, methanol, DMSO, ethylacetate, acetic acid, and ethanolamine.

The terms “compound of this invention,” and “compounds of the presentinvention” and “compounds of Formula I” include compounds of Formulas Iand stereoisomers, geometric isomers, tautomers, solvates, metabolites,and pharmaceutically acceptable salts and prodrugs thereof.

Any formula given herein is intended to represent compounds havingstructures depicted by the structural formula as well as certainvariations or forms. In particular, compounds of any formula givenherein may have asymmetric centers and therefore exist in differentenantiomeric forms. If at least one asymmetrical carbon atom is presentin a compound of the formula I, such a compound may exist in opticallyactive form or in the form of a mixture of optical isomers, e.g. in theform of a racemic mixture. All optical isomers and their mixtures,including the racemic mixtures, are part of the present invention. Thus,any given formula given herein is intended to represent a racemate, oneor more enantiomeric forms, one or more diastereomeric forms, one ormore atropisomeric forms, and mixtures thereof. Certain structures mayexist as geometric isomers (i.e. cis and trans isomers), tautomers, oratropisomers.

Any formula given herein is also intended to represent hydrates,solvates, and polymorphs of such compounds, and mixtures thereof.

Any formula given herein is also intended to represent unlabeled formsas well as isotopically labeled forms of the compounds. Isotopicallylabeled compounds have structures depicted by the formulas given hereinexcept that one or more atoms are replaced by an atom having a selectedatomic mass or mass number. Examples of isotopes that can beincorporated into compounds of the invention include isotopes ofhydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, and chlorine,such as 2H (deuterium), 3H (tritium), 11C, 13C, 14C, 15N, 31P, 32P,.sup.18F 35S, 36Cl, 125I respectively. Various isotopically labeledcompounds of the present invention, for example those into whichradioactive isotopes such as 3H, 13C, and 14C are incorporated. Suchisotopically labelled compounds are useful in metabolic studies,reaction kinetic studies, detection or imaging techniques, such aspositron emission tomography (PET) or single-photon emission computedtomography (SPECT) including drug or substrate tissue distributionassays, or in radioactive treatment of patients. In particular, an 18For labeled compound may be particularly preferred for PET or SPECTstudies. Further, substitution with heavier isotopes such as deuteriummay afford certain therapeutic advantages resulting from greatermetabolic stability, for example increased in vivo half-life or reduceddosage requirements. Isotopically labeled compounds of this inventionand prodrugs thereof can generally be prepared by carrying out theprocedures disclosed in the schemes or in the examples and preparationsdescribed below by substituting a readily available isotopically labeledreagent for a non-isotopically labeled reagent. Further, substitutionwith heavier isotopes, particularly deuterium (i.e., 2H or D) may affordcertain therapeutic advantages resulting from greater metabolicstability, for example increased in vivo half-life or reduced dosagerequirements or an improvement in therapeutic index. It is understoodthat deuterium in this context is regarded as a substituent in thecompound of the formula (I). The concentration of such a heavierisotope, specifically deuterium, may be defined by the isotopicenrichment factor. The term “isotopic enrichment factor” as used hereinmeans the ratio between the isotopic abundance and the natural abundanceof a specified isotope. If a substituent in a compound of this inventionis denoted deuterium, such compound has an isotopic enrichment factorfor each designated deuterium atom of at least 3500 (52.5% deuteriumincorporation at each designated deuterium atom), at least 4000 (60%deuterium incorporation), at least 4500 (67.5% deuterium incorporation),at least 5000 (75% deuterium incorporation), at least 5500 (82.5%deuterium incorporation), at least 6000 (90% deuterium incorporation),at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97%deuterium incorporation), at least 6600 (99% deuterium incorporation),or at least 6633.3 (99.5% deuterium incorporation). In the compounds ofthis invention any atom not specifically designated as a particularisotope is meant to represent any stable isotope of that atom. Unlessotherwise stated, when a position is designated specifically as “H” or“hydrogen”, the position is understood to have hydrogen at its naturalabundance isotopic composition. Accordingly, in the compounds of thisinvention any atom specifically designated as a deuterium (D) is meantto represent deuterium, for example in the ranges given above.

PYRIDO[3,2-d]PYRIMIDINE FORMULA I COMPOUNDS

Formula I compounds include compounds having the formula:

and stereoisomers, geometric isomers, tautomers, or pharmaceuticallyacceptable salts thereof,

wherein

R¹ is selected from H, F, Cl, Br, I, N(R²)₂, OR², SR², SOR², SO₂R²,SO₂N(R²)₂,

C₁-C₁₂ alkyl,

C₂-C₈ alkenyl,

C₂-C₈ alkynyl,

C₆-C₂₀ aryl,

C₃-C₁₂ carbocyclyl,

C₂-C₂₀ heterocyclyl,

C₁-C₂₀ heteroaryl,

—(C₁-C₁₂ alkylene)-(C₃-C₁₂ carbocyclyl),

—(C₁-C₁₂ alkylene)-(C₂-C₂₀ heterocyclyl),

—(C₁-C₁₂ alkylene)-(C₂-C₂₀ heterocyclyl)-(C₂-C₂₀ heterocyclyl),

—(C₁-C₁₂ alkylene)-(C₂-C₂₀ heterocyclyl)-(C₃-C₁₂ carbocyclyl),

—(C₁-C₁₂ alkylene)-(C₂-C₂₀ heterocyclyl)-C(═O)—(C₂-C₂₀ heterocyclyl),

—(C₁-C₁₂ alkylene)-(C₁-C₂₀ heteroaryl),

—(C₁-C₁₂ alkylene)-(C₂-C₂₀ heterocyclyl)-(C₁-C₁₂ alkyl),

—(C₁-C₁₂ alkylene)-(C₆-C₂₀ aryl)-(C₁-C₁₂ alkyl),

—(C₁-C₁₂ alkylene)-(C₁-C₂₀ heteroaryl)-(C₁-C₁₂ alkyl),

—(C₁-C₁₂ alkylene)-C(═O)—(C₂-C₂₀ heterocyclyl),

—(C₁-C₁₂ alkylene)-N(R²)₂,

—(C₁-C₁₂ alkylene)-NR²C(═O)R²,

—(C₁-C₁₂ alkylene)-NR²—(C₁-C₁₂ alkyl),

—(C₁-C₁₂ alkylene)-N(C₁-C₁₂ alkyl)(C₂-C₂₀ heterocyclyl),

—(C₁-C₁₂ alkylene)-NR²—(C₁-C₁₂ alkylene)-(C₁-C₂₀ heteroaryl),

—(C₁-C₁₂ alkylene)-NR²—(C₁-C₁₂ alkylene)-(C₁-C₂₀ heterocyclyl),

—(C₁-C₁₂ alkylene)-NR²—(C₁-C₁₂ alkylene)-NHC(═O)—(C₁-C₂₀ heteroaryl),

—(C₁-C₁₂ alkylene)-(C₂-C₂₀ heterocyclyl)-N(C₁-C₁₂ alkyl)R²,

—(C₁-C₁₂ alkylene)-(C₂-C₂₀ heterocyclyl)-(C₁-C₁₂ alkyl)N(C₁-C₁₂alkyl)R²,

—(C₁-C₁₂ alkylene)-NR²—(C₂-C₂₀ heterocyclyl),

—(C₂-C₁₂ alkenylene)-(C₂-C₂₀ heterocyclyl),

—(C₂-C₂₀ heterocyclyl)-(C₁-C₁₂ alkyl),

—NR²—(C₂-C₂₀ heterocyclyl),

—C(═O)—(C₂-C₂₀ heterocyclyl), and

—C(═O)—(C₁-C₁₂ alkyl),

where alkyl, alkenyl, alkynyl, alkylene, carbocyclyl, heterocyclyl,aryl, and heteroaryl are optionally substituted with one or more groupsindependently selected from F, Cl, Br, I, —CH₃, —CH₂CH₃, —CH₂CH(CH₃)₂,—CH₂OH, —CH₂CH₂OH, —C(CH₃)₂OH, —CH(OH)CH(CH₃)₂, —C(CH₃)₂CH₂OH,—CH₂CH₂SO₂CH₃, —CN, —CF₃, —CHF₂, —CO₂H, —COCH₃, —CO₂CH₃, —C(CH₃)₂CO₂CH₃,—CO₂C(CH₃)₃, —COCH(OH)CH₃, —COCH(CH₃)₂, —CONH₂, —CONHCH₃, —CON(CH₃)₂,—C(CH₃)₂CONH₂, —NO₂, —NH₂, —NHCH₃, —N(CH₃)₂, —NHCOCH₃, —N(CH₃)COCH₃,—NHS(O)₂CH₃, —N(CH₃)C(CH₃)₂CONH₂, —N(CH₃)CH₂CH₂S(O)₂CH₃, ═O, —OH, —OCH₃,—S(O)₂N(CH₃)₂, —SCH₃, —CH₂OCH₃, —S(O)₂CH₃, cyclopropyl, oxetanyl, andmorpholino;

R², R⁴ and R are independently selected from H, C₁-C₁₂ alkyl, C₂-C₈alkenyl, C₂-C₈ alkynyl, —(C₁-C₁₂ alkylene)-(C₃-C₁₂ carbocyclyl),—(C₁-C₁₂ alkylene)-(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂alkylene)-C(═O)—(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂ alkylene)-(C₆-C₂₀ aryl),and —(C₁-C₁₂ alkylene)-(C₁-C₂₀ heteroaryl), where alkyl, alkenyl,alkynyl, alkylene, carbocyclyl, heterocyclyl, aryl, and heteroaryl areoptionally substituted with one or more groups independently selectedfrom F, Cl, Br, I, —CH₃, —CH₂OH, —CN, —CF₃, —CO₂H, —COCH₃, —COC(CH₃)₃,—COCF₃, —CH₂CF₃, —CO₂CH₃, —CONH₂, —CONHCH₃, —CON(CH₃)₂, —NO₂, —NH₂,—NHCH₃, —NHCOCH₃, —NHS(O)₂CH₃, —OH, —OCH₃, —OCH₂CH₃, —OCH(CH₃)₂,—S(O)₂N(CH₃)₂, —SCH₃, —CH₂OCH₃, and —S(O)₂CH₃;

R³ is selected from C₆-C₂₀ aryl, C₂-C₂₀ heterocyclyl and C₁-C₂₀heteroaryl, each of which are optionally substituted with one or moregroups independently selected from F, Cl, Br, I, —CH₃, —CH₂CH₃,—CH(CH₃)₂, —CH₂CN, —CN, —CHF₂, —CF₃, —CH₂CF₃, —CF₂CH₃, —CH₂OH,—CH₂CH₂OH, —CH(CH₃)OH, —CH(CH₃)OCH₃, —CO₂H, —CONH₂, —CON(CH₃)₂, —NO₂,—NH₂, —NHCH₃, —N(CH₃)₂, —NHCOCH₃, —OH, —OCH₃, —SH, —NHC(═O)NHCH₃,—NHC(═O)NHCH₂CH₃, —S(O)₂CH₃, cyclopropyl, pyrrolidin-1-yl,3-methoxyazetidin-1-yl, and azetidin-1-yl;

R⁵ is selected from H, C₁-C₁₂ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl,—(C₁-C₁₂ alkylene)-(C₃-C₁₂ carbocyclyl), —(C₁-C₁₂ alkylene)-(C₂-C₂₀heterocyclyl), —(C₁-C₁₂ alkylene)-C(═O)—(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂alkylene)-(C₆-C₂₀ aryl), and —(C₁-C₁₂ alkylene)-(C₁-C₂₀ heteroaryl),where alkyl, alkenyl, alkynyl, alkylene, carbocyclyl, heterocyclyl,aryl, and heteroaryl are optionally substituted with one or more groupsindependently selected from F, Cl, Br, I, —CH₃, —CH₂OH, —CN, —CF₃,—CO₂H, —COCH₃, —CO₂CH₃, —CONH₂, —CONHCH₃, —CON(CH₃)₂, —NO₂, —NH₂,—NHCH₃, —NHCOCH₃, —NHS(O)₂CH₃, —OH, —OCH₃, —S(O)₂N(CH₃)₂, —SCH₃,—CH₂OCH₃, and —S(O)₂CH₃;

and n is 0 or 1.

Further it is to be understood that every embodiment relating to aspecific residue R¹, R², R³, R⁴ and R⁵ as disclosed herein may becombined with any other embodiment relating to another residue R¹, R²,R³, R⁴ and R⁵ as disclosed herein.

Exemplary embodiments of Formula I compounds include wherein R¹ isselected from the structures

where the wavy line indicates the site of attachment.

Exemplary embodiments of Formula I compounds include wherein R² or R⁴are independently C₁-C₁₂ alkyl optionally substituted with one or moregroups independently selected from F, Cl, Br, I, —CH₃, —CH₂OH, —CN,—CF₃, —CO₂H, —COCH₃, —CO₂CH₃, —CONH₂, —CONHCH₃, —CON(CH₃)₂, —NO₂, —NH₂,—NHCH₃, —NHCOCH₃, —NHS(O)₂CH₃, —OH, —OCH₃, —S(O)₂N(CH₃)₂, —SCH₃,—CH₂OCH₃, and —S(O)₂CH₃.

Exemplary embodiments of Formula I compounds include wherein R² or R⁴ isindependently CH₃, or R² and R⁴ are each H.

Exemplary embodiments of Formula I compounds include wherein R³ isselected from:

each of which are optionally substituted with one or more groupsindependently selected from F, Cl, Br, I, —CH₃, —CH₂CH₃, —CH₂CH₂CH₃,—CH(CH₃)₂, —C(CH₃)₃, —CH₂OCH₃, —CHF₂, —CN, —CF₃, —CH₂OH, —CH₂OCH₃,—CH₂CH₂OH, —CH₂C(CH₃)₂OH, —CH(CH₃)OH, —CH(CH₂CH₃)OH, —CH₂CH(OH)CH₃,—CH₂CH(OCH₃)CH₃, —C(CH₃)₂OH, —C(CH₃)₂OCH₃, —CH(CH₃)F, —C(CH₃)F₂,—CH(CH₂CH₃)F, —C(CH₂CH₃)₂F, —CO₂H, —CONH₂, —CON(CH₂CH₃)₂, —COCH₃,—CON(CH₃)₂, —NO₂, —NH₂, —NHCH₃, —N(CH₃)₂, —NHCH₂CH₃, —NHCH(CH₃)₂,—NHCH₂CH₂OH, —NHCH₂CH₂OCH₃, —NHCOCH₃, —NHCOCH₂CH₃, —NHCOCH₂OH,—NHS(O)₂CH₃, —N(CH₃)S(O)₂CH₃, ═O, —OH, —OCH₃, —OCH₂CH₃, —OCH(CH₃)₂, —SH,—NHC(═O)NHCH₃, —NHC(═O)NHCH₂CH₃, —S(O)CH₃, —S(O)CH₂CH₃, —S(O)₂CH₃,—S(O)₂NH₂, —S(O)₂NHCH₃, —S(O)₂N(CH₃)₂, —CH₂S(O)₂CH₃, and a groupselected from

Exemplary embodiments of Formula I compounds include wherein R³ is amonocyclic heteroaryl selected from pyridyl, isoxazolyl, imidazolyl,pyrazolyl, pyrrolyl, thiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl,oxazolyl, oxadiazolyl, furanyl, thienyl, triazolyl, and tetrazolyl.

Exemplary embodiments of Formula I compounds include wherein R³ is amonocyclic heteroaryl selected from:

where the wavy line indicates the site of attachment.

Exemplary embodiments of Formula I compounds include wherein R³ is amonocyclic heteroaryl selected from:

where the wavy line indicates the site of attachment.

Exemplary embodiments of Formula I compounds include wherein R³ is amonocyclic heteroaryl selected from:

where the wavy line indicates the site of attachment.

Exemplary embodiments of Formula I compounds include wherein R³ is acarbon-linked, fused bicyclic C₄-C₂₀ heterocyclyl or C₁-C₂₀ heteroarylselected from

where the wavy line indicates the site of attachment.

Exemplary embodiments of Formula I compounds include wherein R³ isselected from:

where the wavy line indicates the site of attachment and R¹⁴ is selectedfrom F, Cl, Br, I, —CH₃, —CN, —CF₃, —CH₂OH, —CO₂H, —CONH₂, —CON(CH₃)₂,—NO₂, —NH₂, —NHCH₃, —NHCOCH₃, —OH, —OCH₃, —SH, —NHC(═O)NHCH₃,—NHC(═O)NHCH₂CH₃, and —S(O)₂CH₃.

Exemplary embodiments of Formula I compounds include wherein R³ is acarbon-linked, fused bicyclic C₄-C₂₀ heterocyclyl or C₁-C₂₀ heteroarylselected from:

where the wavy line indicates the site of attachment.

Exemplary embodiments of Formula I compounds include wherein R³ is1H-indazol-4-yl and n is 0.

Alternatively, Formula I compounds include compounds having the formula:

and stereoisomers, geometric isomers, tautomers, or pharmaceuticallyacceptable salts thereof,

wherein

R¹ is selected from C₁-C₁₂ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₆-C₂₀aryl, C₃-C₁₂ carbocyclyl, C₂-C₂₀ heterocyclyl, C₁-C₂₀ heteroaryl,—(C₁-C₁₂ alkylene)-(C₃-C₁₂ carbocyclyl), —(C₁-C₁₂ alkylene)-(C₂-C₂₀heterocyclyl), —(C₁-C₁₂ alkylene)-(C₂-C₂₀ heterocyclyl)-(C₂-C₂₀heterocyclyl), —(C₁-C₁₂ alkylene)-(C₂-C₂₀ heterocyclyl)-(C₃-C₁₂carbocyclyl), —(C₁-C₁₂ alkylene)-(C₂-C₂₀ heterocyclyl)-C(═O)—(C₂-C₂₀heterocyclyl), —(C₁-C₁₂ alkylene)-(C₁-C₂₀ heteroaryl), —(C₁-C₁₂alkylene)-(C₂-C₂₀ heterocyclyl)-(C₁-C₁₂ alkyl), —(C₁-C₁₂alkylene)-(C₆-C₂₀ aryl)-(C₁-C₁₂ alkyl), —(C₁-C₁₂ alkylene)-(C₁-C₂₀heteroaryl)-(C₁-C₁₂ alkyl), —(C₁-C₁₂ alkylene)-C(═O)—(C₂-C₂₀heterocyclyl), —(C₁-C₁₂ alkylene)-NHR², —(C₁-C₁₂ alkylene)-NR²—(C₁-C₁₂alkyl), —(C₁-C₁₂ alkylene)-N(C₁-C₁₂ alkyl)(C₂-C₂₀ heterocyclyl),—(C₁-C₁₂ alkylene)-NR²—(C₁-C₁₂ alkylene)-(C₁-C₂₀ heteroaryl), —(C₁-C₁₂alkylene)-NR²—(C₁-C₁₂ alkylene)-(C₁-C₂₀ heterocyclyl), —(C₁-C₁₂alkylene)-NR²—(C₁-C₁₂ alkylene)-NHC(═O)—(C₁-C₂₀ heteroaryl), —(C₁-C₁₂alkylene)-(C₂-C₂₀ heterocyclyl)-N(C₁-C₁₂ alkyl)R², —(C₁-C₁₂alkylene)-(C₂-C₂₀ heterocyclyl)-(C₁-C₁₂ alkyl)-N(C₁-C₁₂ alkyl)R²,—(C₁-C₁₂ alkylene)-NR²—(C₂-C₂₀ heterocyclyl), —(C₂-C₁₂alkenylene)-(C₂-C₂₀ heterocyclyl), —NR²—(C₂-C₂₀ heterocyclyl),—C(═O)-(C₂-C₂₀ heterocyclyl), and —C(═O)—(C₁-C₁₂ alkyl), where alkyl,alkenyl, alkynyl, alkylene, carbocyclyl, heterocyclyl, aryl, andheteroaryl are optionally substituted with one or more groupsindependently selected from F, Cl, Br, I, —CH₃, —CH₂CH₃, —CH₂CH(CH₃)₂,—CH₂OH, —CH₂CH₂OH, —C(CH₃)₂OH, —CH(OH)CH(CH₃)₂, —C(CH₃)₂CH₂OH,—CH₂CH₂SO₂CH₃, —CN, —CF₃₅-CO₂H, —COCH₃, —CO₂CH₃, —CO₂C(CH₃)₃,—COCH(OH)CH₃, —CONH₂, —CONHCH₃, —CON(CH₃)₂, —C(CH₃)₂CONH₂, —NO₂, —NH₂,—NHCH₃, —N(CH₃)₂, —NHCOCH₃, —N(CH₃)COCH₃, —NHS(O)₂CH₃,—N(CH₃)C(CH₃)₂CONH₂, —N(CH₃)CH₂CH₂S(O)₂CH₃, ═O, —OH, —OCH₃,—S(O)₂N(CH₃)₂, —SCH₃, —CH₂OCH₃, —S(O)₂CH₃, cyclopropyl, oxetanyl, andmorpholino;

R² and R⁴ are independently selected from H, C₁-C₁₂ alkyl, C₂-C₈alkenyl, C₂-C₈ alkynyl, —(C₁-C₁₂ alkylene)-(C₃-C₁₂ carbocyclyl),—(C₁-C₁₂ alkylene)-(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂alkylene)-C(═O)—(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂ alkylene)-(C₆-C₂₀ aryl),and —(C₁-C₁₂ alkylene)-(C₁-C₂₀ heteroaryl), where alkyl, alkenyl,alkynyl, alkylene, carbocyclyl, heterocyclyl, aryl, and heteroaryl areoptionally substituted with one or more groups independently selectedfrom F, Cl, Br, I, —CH₃, —CH₂OH, —CN, —CF₃, —CO₂H, —COCH₃, —COC(CH₃)₃,—COCF₃, —CO₂CH₃, —CONH₂, —CONHCH₃, —CON(CH₃)₂, —NO₂, —NH₂, —NHCH₃,—NHCOCH₃, —NHS(O)₂CH₃, —OH, —OCH₃, —S(O)₂N(CH₃)₂, —SCH₃, —CH₂OCH₃, and—S(O)₂CH₃;

R³ is selected from C₆-C₂₀ aryl, C₂-C₂₀ heterocyclyl and C₁-C₂₀heteroaryl, each of which are optionally substituted with one or moregroups independently selected from F, Cl, Br, I, —CH₃, —CN, —CF₃,—CH₂OH, —CO₂H, —CONH₂, —CON(CH₃)₂, —NO₂, —NH₂, —NHCH₃, —NHCOCH₃, —OH,—OCH₃, —SH, —NHC(═O)NHCH₃, —NHC(═O)NHCH₂CH₃, and —S(O)₂CH₃;

R⁵ is selected from H, C₁-C₁₂ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl,—(C₁-C₁₂ alkylene)-(C₃-C₁₂ carbocyclyl), —(C₁-C₁₂ alkylene)-(C₂-C₂₀heterocyclyl), —(C₁-C₁₂ alkylene)-C(═O)—(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂alkylene)-(C₆-C₂₀ aryl), and —(C₁-C₁₂ alkylene)-(C₁-C₂₀ heteroaryl),where alkyl, alkenyl, alkynyl, alkylene, carbocyclyl, heterocyclyl,aryl, and heteroaryl are optionally substituted with one or more groupsindependently selected from F, Cl, Br, I, —CH₃, —CH₂OH, —CN, —CF₃,—CO₂H, —COCH₃, —CO₂CH₃, —CONH₂, —CONHCH₃, —CON(CH₃)₂, —NO₂, —NH₂,—NHCH₃, —NHCOCH₃, —NHS(O)₂CH₃, —OH, —OCH₃, —S(O)₂N(CH₃)₂, —SCH₃,—CH₂OCH₃, and —S(O)₂CH₃;

and n is 0 or 1.

The Formula I compounds of the invention may contain asymmetric orchiral centers, and therefore exist in different stereoisomeric forms.It is intended that all stereoisomeric forms of the compounds of theinvention, including but not limited to, diastereomers, enantiomers andatropisomers, as well as mixtures thereof such as racemic mixtures, formpart of the present invention.

In addition, the present invention embraces all geometric and positionalisomers. For example, if a Formula I compound incorporates a double bondor a fused ring, the cis- and trans-forms, as well as mixtures thereof,are embraced within the scope of the invention. Both the singlepositional isomers and mixture of positional isomers are also within thescope of the present invention.

In the structures shown herein, where the stereochemistry of anyparticular chiral atom is not specified, then all stereoisomers arecontemplated and included as the compounds of the invention. Wherestereochemistry is specified by a solid wedge or dashed linerepresenting a particular configuration, then that stereoisomer is sospecified and defined.

The compounds of the present invention may exist in unsolvated as wellas solvated forms with pharmaceutically acceptable solvents such aswater, ethanol, and the like, and it is intended that the inventionembrace both solvated and unsolvated forms.

The compounds of the present invention may also exist in differenttautomeric forms, and all such forms are embraced within the scope ofthe invention. The term “tautomer” or “tautomeric form” refers tostructural isomers of different energies which are interconvertible viaa low energy barrier. For example, proton tautomers (also known asprototropic tautomers) include interconversions via migration of aproton, such as keto-enol and imine-enamine isomerizations. Valencetautomers include interconversions by reorganization of some of thebonding electrons.

The present invention also embraces isotopically-labeled compounds ofthe present invention which are identical to those recited herein, butfor the fact that one or more atoms are replaced by an atom having anatomic mass or mass number different from the atomic mass or mass numberusually found in nature. All isotopes of any particular atom or elementas specified are contemplated within the scope of the compounds of theinvention, and their uses. Exemplary isotopes that can be incorporatedinto compounds of the invention include isotopes of hydrogen, carbon,nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine and iodine,such as ²H, ³H, ¹¹C, ¹⁴C, ¹³N, ¹⁵O, ¹⁷O, ¹⁸O, ³²P, ³⁵S, ¹⁸F, ³⁶Cl, ¹²³Iand ¹²⁵I. Certain isotopically-labeled compounds of the presentinvention (e.g., those labeled with ³H and ¹⁴C) are useful in compoundand/or substrate tissue distribution assays. Tritiated (³H) andcarbon-14 (⁴C) isotopes are useful for their ease of preparation anddetectability. Further, substitution with heavier isotopes such asdeuterium (i.e., ²H) may afford certain therapeutic advantages resultingfrom greater metabolic stability (e.g., increased in vivo half-life orreduced dosage requirements) and hence may be preferred in somecircumstances. Positron emitting isotopes such as ¹⁵O, ¹³N, ¹¹C and ¹⁸Fare useful for positron emission tomography (PET) studies to examinesubstrate receptor occupancy. Isotopically labeled compounds of thepresent invention can generally be prepared by following proceduresanalogous to those disclosed in the Schemes and/or in the Examplesherein below, by substituting an isotopically labeled reagent for anon-isotopically labeled reagent.

Biological Evaluation

The relative efficacies of Formula I compounds as inhibitors of anenzyme activity (or other biological activity) can be established bydetermining the concentrations at which each compound inhibits theactivity to a predefined extent and then comparing the results.Typically, the preferred determination is the concentration thatinhibits 50% of the activity in a biochemical assay, i.e., the 50%inhibitory concentration or “IC₅₀”. Determination of IC₅₀ values can beaccomplished using conventional techniques known in the art. In general,an IC₅₀ can be determined by measuring the activity of a given enzyme inthe presence of a range of concentrations of the inhibitor under study.The experimentally obtained values of enzyme activity then are plottedagainst the inhibitor concentrations used. The concentration of theinhibitor that shows 50% enzyme activity (as compared to the activity inthe absence of any inhibitor) is taken as the IC₅₀ value. Analogously,other inhibitory concentrations can be defined through appropriatedeterminations of activity. For example, in some settings it can bedesirable to establish a 90% inhibitory concentration, i.e., IC₉₀, etc.

Accordingly, a “selective PI3K delta inhibitor” can be understood torefer to a compound that exhibits a 50% inhibitory concentration (IC₅₀)with respect to PI3K delta that is at least at least 10-fold lower thanthe IC₅₀ value with respect to any or all of the other Class I PI3Kfamily members.

Determination of the activity of PI3 kinase activity of Formula Icompounds is possible by a number of direct and indirect detectionmethods. Certain exemplary compounds described herein were assayed fortheir ability to inhibit PI3K alpha, beta, gamma, and delta isoforms(Example 901). The range of IC₅₀ values for inhibition of PI3K delta wasless than 1 nM (nanomolar) to about 10 μM (micromolar). Certainexemplary compounds of the invention had PI3K delta inhibitory IC₅₀values less than 10 nM. The compounds are selective for the p110δ(delta) isoform, which is a class Ia PI3 kinase, over other class Ia PI3kinases, and are thus selective for the p110δ isoform over both thep110α (alpha) isoform and the p110β (beta) isoform. In particular, theyare selective for p110δ (delta) over p110α (alpha). The compounds arealso selective for the p110δ isoform over p110γ (gamma), which is aclass Ib kinase. The selectivity exhibited by certain Formula Icompounds of the invention for p110δ (delta) over the p110α (alpha)isoform of PI3 kinase is at least 10 fold, as exemplified by the ratiosof biochemical IC₅₀ values (Example 901).

Certain Formula I compounds may have antiproliferative activity to treathyperproliferative disorders such as cancer. The Formula I compounds mayinhibit tumor growth in mammals and may be useful for treating humancancer patients. Formula I compounds may be tested for in vitro cellproliferation activity and in vivo tumor growth inhibition according tothe methods in WO 2006/046031; US 2008/0039459; US 2008/0076768; US2008/0076758; WO 2008/070740; WO 2008/073785, which are incorporated byreference herein.

Evaluation of drug-induced immunosuppression by the compounds of theinvention may be performed using in vivo functional tests, such asrodent models of induced arthritis and therapeutic or prophylactictreatment to assess disease score, T cell-dependent antibody response(TDAR), and delayed-type hypersensitivity (DTH). Other in vivo systemsincluding murine models of host defense against infections or tumorresistance (Burleson G R, Dean J H, and Munson A E. Methods inImmunotoxicology, Vol. 1. Wiley-Liss, New York, 1995) may be consideredto elucidate the nature or mechanisms of observed immunosuppression. Thein vivo test systems can be complemented by well-established in vitro orex vivo functional assays for the assessment of immune competence. Theseassays may comprise B or T cell proliferation in response to mitogens orspecific antigens, measurement of signaling through the PI3K pathway inB or T cells or immortalized B or T cell lines, measurement of cellsurface markers in response to B or T cell signaling, natural killer(NK) cell activity, mast cell activity, mast cell degranulation,macrophage phagocytosis or kill activity, and neutrophil oxidative burstand/or chemotaxis. In each of these tests determination of cytokineproduction by particular effector cells (e.g., lymphocytes, NK,monocytes/macrophages, neutrophils) may be included. The in vitro and exvivo assays can be applied in both preclinical and clinical testingusing lymphoid tissues and/or peripheral blood (House R V. “Theory andpractice of cytokine assessment in immunotoxicology” (1999) Methods19:17-27; Hubbard A K. “Effects of xenobiotics on macrophage function:evaluation in vitro” (1999) Methods; 19:8-16; Lebrec H, et al (2001)Toxicology 158:25-29).

Collagen-Induced Arthritis (CIA) 6-week detailed study using anautoimmune mechanism to mimic human arthritis; rat and mouse models(Example 902). Collagen-induced arthritis (CIA) is one of the mostcommonly used animal models of human rheumatoid arthritis (RN). Jointinflammation, which develops in animals with CIA, strongly resemblesinflammation observed in patients with RA. Blocking tumor necrosisfactor (TNF) is an efficacious treatment of CIA, just as it is a highlyefficacious therapy in treatment of RA patients. CIA is mediated by bothT-cells and antibodies (B-cells). Macrophages are believed to play animportant role in mediating tissue damage during disease development.CIA is induced by immunizing animals with collagen emulsified inComplete Freund's Adjuvant (CFA). It is most commonly induced in theDBA/1 mouse strain, but the disease can also be induced in Lewis rats.

There is good evidence that B-cells play a key role in the pathogenesisof autoimmune and/or inflammatory disease. Protein-based therapeuticsthat deplete B cells such as Rituxan are effective againstautoantibody-driven inflammatory diseases such as rheumatoid arthritis(Rastetter et al. (2004) Annu Rev Med 55:477). CD69 is the earlyactivation marker in leukocytes including T cells, thymocytes, B cells,NK cells, neutrophils, and eosinophils. The CD69 human whole blood assay(Example 903) determines the ability of compounds to inhibit theproduction of CD69 by B lymphocytes in human whole blood activated bycrosslinking surface IgM with goat F(ab′)2 anti-human IgM.

The T-cell Dependent Antibody Response (TDAR) is a predictive assay forimmune function testing when potential immunotoxic effects of compoundsneed to be studied. The Levi-Plaque Forming Cell (PFC) assay, usingSheep Red Blood Cells (SRBC) as the antigen, is currently a widelyaccepted and validated standard test. TDAR has proven to be a highlypredictable assay for adult exposure immunotoxicity detection in micebased on the US National Toxicology Program (NTP) database (M. I. Lusteret al (1992) Fundam. Appl. Toxicol. 18:200-210). The utility of thisassay stems from the fact that it is a holistic measurement involvingseveral important components of an immune response. A TDAR is dependenton functions of the following cellular compartments: (1)antigen-presenting cells, such as macrophages or dendritic cells; (2)T-helper cells, which are critical players in the genesis of theresponse, as well as in isotype switching; and (3) B-cells, which arethe ultimate effector cells and are responsible for antibody production.Chemically-induced changes in any one compartment can cause significantchanges in the overall TDAR (M. P. Holsapple In: G. R. Burleson, J. H.Dean and A. E. Munson, Editors, Modern Methods in Immunotoxicology,Volume 1, Wiley-Liss Publishers, New York, N.Y. (1995), pp. 71-108).Usually, this assay is performed either as an ELISA for measurement ofsoluble antibody (R. J. Smialowizc et al (2001) Toxicol. Sci.61:164-175) or as a plaque (or antibody) forming cell assay (L. Guo etal (2002) Toxicol. Appl. Pharmacol. 181:219-227) to detect plasma cellssecreting antigen specific antibodies. The antigen of choice is eitherwhole cells (e.g. sheep erythrocytes) or soluble protein antigens (T.Miller et al (1998) Toxicol. Sci. 42:129-135).

Exemplary Formula I compounds No. 101-201 in Tables 1 and 2 were made,characterized, and tested for inhibition of PI3K delta and selectivityaccording to the methods of this invention, and have the followingstructures and corresponding names (ChemDraw Ultra, Version 9.0.1,CambridgeSoft Corp., Cambridge Mass.).

TABLE 1 No. Structure Name 101

2-(1-((2-(2-methyl-1H- benzo[d]imidazol-1-yl)-4- morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-4- yl)propan-2-ol 102

2-(1-((2-(2-cyclopropyl-1H- benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2- d]pyrimidin-6- yl)methyl)piperidin-4-yl)propan-2-ol 103

2-(1-((2-(2-methylbenzofuran- 3-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-4- yl)propan-2-ol 104

2-ethyl-1-(6-((4-(2- hydroxypropan-2-yl)piperidin- 1-yl)methyl)-4-morpholinopyrido[3,2- d]pyrimidin-2-yl)-1H-indazol- 3(2H)-one 105

2-(1-((2-(5-fluoro-1H-indol-4- yl)-4-morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-4- yl)propan-2-ol 106

2-(1-((2-(5-methyl-1H- pyrazol-3-ylamino)-4- morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-4- yl)propan-2-ol 107

2-(1-((2-(2-aminopyrimidin-5- yl)-4-morpholinopyrido[3,2- d]pyrimidin-6-yl)methyl)piperidin-4- yl)propan-2-ol 108

2-(5-fluoro-1H-indol-4-yl)-4- morpholinopyrido[3,2-d]pyrimidin-6-yl)methanol 109

2-(1-((2-(1H-indol-4-yl)-4- morpholinopyrido[3,2- d]pyrimidin-6-yl)methyl)piperidin-4- yl)propan-2-ol 110

2-(1-((2-(1H-indazol-4-yl)-4- morpholinopyrido[3,2- d]pyrimidin-6-yl)methyl)piperidin-4- yl)propan-2-ol 111

4-(2-(5-fluoro-1H-indol-4-yl)- 6-((3-(tetrahydro-2H-pyran-4-yl)azetidin-1- yl)methyl)pyrido[3,2- d]pyrimidin-4-yl)morpholine 112

2-(1-((2-(2-isopropyl-1H- benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2- d]pyrimidin-6- yl)methyl)piperidin-4-yl)propan-2-ol 113

4-(2-(2-isopropyl-1H- benzo[d]imidazol-1-yl)-6-((3-(tetrahydro-2H-pyran-4- yl)azetidin-1- yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine 114

(S)-2-(1-((2-(2-(1- methoxyethyl)-1H- benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2- d]pyrimidin-6- yl)methyl)piperidin-4-yl)propan-2-ol 115

(R)-2-(1-((2-(2-(1- methoxyethyl)-1H- benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2- d]pyrimidin-6- yl)methyl)piperidin-4-yl)propan-2-ol 116

2-(1-((2-(6-amino-2- methylpyridin-3-yl)-4- morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-4- yl)propan-2-ol 117

methyl 2-(3-((2-(2-isopropyl- 1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2- d]pyrimidin-6- yl)methyl)azetidin-1-yl)-2-methylpropanoate 118

(2-(5-fluoro-1H-indol-4-yl)-4- morpholinopyrido[3,2-d]pyrimidin-6-yl)(4-(2- hydroxypropan-2-yl)piperidin- 1-yl)methanone 119

(4-(2-hydroxypropan-2- yl)piperidin-1-yl)(2-(2- isopropyl-1H-benzo[d]imidazol-1-yl)-4- morpholinopyrido[3,2-d]pyrimidin-6-yl)methanone 120

2-(1-((2-(2-(1,1- difluoroethyl)-1H- benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2- d]pyrimidin-6- yl)methyl)piperidin-4-yl)propan-2-ol 121

2-(4-((2-(5-fluoro-1H-indol-4- yl)-4-morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)piperazin-1-yl)-2- methylpropanamide 122

2-(1-((2-(6-aminopyridin-3- yl)-4-morpholinopyrido[3,2- d]pyrimidin-6-yl)methyl)piperidin-4- yl)propan-2-ol 123

2-(4-((2-(1H-indol-3-yl)-4- morpholinopyrido[3,2- d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2- methylpropanamide 124

2-(4-((2-(6-aminopyridin-3- yl)-4-morpholinopyrido[3,2- d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2- methylpropanamide 125

4-(6-((3-(4,4- difluoropiperidin-1- yl)azetidin-1-yl)methyl)-2-(5-fluoro-1H-indol-4- yl)pyrido[3,2-d]pyrimidin-4- yl)morpholine 126

4-(1-((2-(5-fluoro-1H-indol-4- yl)-4-morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)azetidin-3- yl)morpholine 127

4-(1-((2-(5-fluoro-1H-indol-4- yl)-4-morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)azetidin-3- yl)piperazin-2-one 128

2-(1-((2-(1H-indol-1-yl)-4- morpholinopyrido[3,2- d]pyrimidin-6-yl)methyl)piperidin-4- yl)propan-2-ol 129

4-(2-(5-fluoro-1H-indol-4-yl)- 6-((3-(1,1- dioxo)thiomorpholinoazetidin-1-yl)methyl)pyrido[3,2- d]pyrimidin-4-yl)morpholine 130

4-(2-(5-fluoro-1H-indol-4-yl)- 6-((4-(oxetan-3-yl)piperidin-1-yl)methyl)pyrido[3,2- d]pyrimidin-4-yl)morpholine 131

4-(1-((2-(2-ethyl-1H- benzo[d]imidazol-1-yl)-4- morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)azetidin-3- yl)mopholine 132

2-(1-((2-(6-amino-5- methylpyridin-3-yl)-4- morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-4- yl)propan-2-ol 133

2-(1-((2-(1H-indol-3-yl)-4- morpholinopyrido[3,2- d]pyrimidin-6-yl)methyl)piperidin-4- yl)propan-2-ol 134

4-(1-((2-(2-ethyl-1H- benzo[d]imidazol-1-yl)-4- morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)azetidin-3- yl)piperazin-2-one

TABLE 2 No. Structure Name 135

2-(1-((2-(2-methyl-1H-indol- 1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-4- yl)propan-2-ol 136

2-(1-((2-(2-ethyl-2H-indazol- 3-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-4- yl)propan-2-ol 137

tert-butyl 4-((2-(5-fluoro-1H- indol-4-yl)-4- morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)piperidine-1- carboxylate 138

2-(1-((2-(1H-indazol-3-yl)-4- morpholinopyrido[3,2- d]pyrimidin-6-yl)methyl)piperidin-4- yl)propan-2-ol 139

4-(2-(5-fluoro-1H-indol-4-yl)- 6-(piperidin-4- ylmethyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine 140

1-(((2-(5-fluoro-1H-indol-4- yl)-4-morpholinopyrido[3,2- d]pyrimidin-6-yl)methyl)(methyl)amino)-2- methylpropan-2-ol 141

4-(2-(5-fluoro-1H-indol-4-yl)- 6-(1-isopropyl-1H-1,2,4-triazol-5-yl)pyrido[3,2- d]pyrimidin-4-yl)morpholine 142

N-((2-(5-fluoro-1H-indol-4- yl)-4-morpholinopyrido[3,2- d]pyrimidin-6-yl)methyl)tetrahydro-2H- pyran-4-amine 143

1-((2-(5-fluoro-1H-indol-4-yl)- 4-morpholinopyrido[3,2- d]pyrimidin-6-yl)methylamino)-2- methylpropan-2-ol 144

4-(6-((3,3-dimethylpyrrolidin- 1-yl)methyl)-2-(5-fluoro-1H-indol-4-yl)pyrido[3,2- d]pyrimidin-4-yl)morpholine 145

N-((2-(5-fluoro-1H-indol-4- yl)-4-morpholinopyrido[3,2- d]pyrimidin-6-yl)methyl)pivalamide 146

4-((2-(5-fluoro-1H-indol-4-yl)- 4-morpholinopyrido[3,2- d]pyrimidin-6-yl)methyl)morpholine 147

N-((2-(5-fluoro-1H-indol-4- yl)-4-morpholinopyrido[3,2- d]pyrimidin-6-yl)methyl)isobutyramide 148

1-(4-((2-(5-fluoro-1H-indol-4- yl)-4-morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-1- yl)ethanone 149

1-(4-((2-(5-fluoro-1H-indol-4- yl)-4-morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-1-yl)-2- methylpropan-1-one 150

(S)-4-(1-((2-(2-(1- methoxyethyl)-1H- benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2- d]pyrimidin-6- yl)methyl)azetidin-3- yl)morpholine151

2-(2-(2-aminopyrimidin-5-yl)- 4-morpholinopyrido[3,2-d]pyrimidin-6-yl)propan-2-ol 152

5-(6-(2-methoxypropan-2-yl)- 4-morpholinopyrido[3,2-d]pyrimidin-2-yl)pyrimidin-2- amine 153

2-(1-((2-(2-(dimethylamino)- 1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2- d]pyrimidin-6- yl)methyl)piperidin-4-yl)propan-2-ol 154

4-(2-(5-fluoro-1H-indol-4-yl)- 6-((tetrahydro-2H-pyran-4-yl)methyl)pyrido[3,2- d]pyrimidin-4-yl)morpholine 155

5-(6-(difluoromethyl)-4- morpholinopyrido[3,2- d]pyrimidin-2-yl)-4-methylpyrimidin-2-amine 156

2-(1-((4-morpholino-2-(2- (trifluoromethyl)-1H- benzo[d]imidazol-1-yl)pyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-4- yl)propan-2-ol 157

2-(1-((2-(2-(difluoromethyl)- 1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2- d]pyrimidin-6- yl)methyl)piperidin-4-yl)propan-2-ol 158

4-(1-(6-((4-(2-hydroxypropan- 2-yl)piperidin-1-yl)methyl)-4-morpholinopyrido[3,2- d]pyrimidin-2-yl)-1H- benzo[d]imidazol-2-yl)acetonitrile 159

4,4′-(6-(5-fluoro-1H-indol-4- yl)pyrido[3,2-d]pyrimidine-2,4-diyl)dimorpholine 160

2-(1-((2-(2- (methylamino)pyridin-3-yl)-4- morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-4- yl)propan-2-ol 161

2-(1-((2-(imidazo[1,2- a]pyridin-5-yl)-4- morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-4- yl)propan-2-ol 162

5-(6-(difluoromethyl)-4- morpholinopyrido[3,2-d]pyrimidin-2-yl)pyrimidin-2- amine 163

N,N-dimethyl-1-(4- morpholino-6-((3-(1,1- dioxo)thiomorpholinoazetidin-1-yl)methyl)pyrido[3,2- d]pyrimidin-2-yl)-1H- benzo[d]imidazol-2-amine164

(S)-1-(4-((2-(2-(1- methoxyethyl)-1H- benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2- d]pyrimidin-6- yl)methyl)piperidin-1- yl)ethanone165

1-(4-((2-(2-(dimethylamino)- 1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2- d]pyrimidin-6- yl)methyl)piperidin-1- yl)ethanone166

2-(1-((2-([1,2,4]triazolo[1,5- a]pyridin-5-yl)-4- morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-4- yl)propan-2-ol 167

N,N-dimethyl-1-(4- morpholino-6-((3- morpholinoazetidin-1-yl)methyl)pyrido[3,2- d]pyrimidin-2-yl)-1H- benzo[d]imidazol-2-amine 168

2-(1-((2-(2-(2-hydroxyethyl)- 1H-benzo[d]imidazol-1-yl)-4-mropholinopyrido[3,2- d]pyrimidin-6- yl)methyl)piperidin-4-yl)propan-2-ol 169

4-(2-(2-(difluoromethyl)-1H- benzo[d]imidazol-1-yl)-6-((3- (1,1-dioxo)thiomorpholinoazetidin- 1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine 170

2-(1-((2-(2-methoxy-1H- benzo[d]imidazol-1-yl)-4- morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-4- yl)propan-2-ol 171

1-(4-((2-(2-(difluoromethyl)- 1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2- d]pyrimidin-6- yl)methyl)piperidin-1- yl)ethanone172

4-(1-((2-(2-(difluoromethyl)- 1H-benzo[d]imidazol-1-yl)-4-morpholinepyrido[3,2- d]pyrimidin-6- yl)methyl)azetidin-3- yl)morpholine173

2-(1-((4-morpholino-2-(2- (2,2,2-trifluoroethyl)-1H- benzo[d]imidazol-1-yl)pyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-4- yl)propan-2-ol 174

5-(4-morpholinopyrido[3,2- d]pyrimidin-2-yl)pyrimidin-2- amine 175

5-(6-methoxy-4- morpholinopyrido[3,2- d]pyrimidin-2-yl)pyrimidin-2-amine 176

2-(1-((2-(2-(azetidin-1-yl)-1H- benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2- d]pyrimidin-6- yl)methyl)piperidin-4-yl)propan-2-ol 177

2-(1-((4-morpholino-2-(2- (pyrrolidin-1-yl)-1H- benzo[d]imidazol-1-yl)pyrido[3,2-d]pyrimidin-6- yl)methyl)piperidin-4- yl)propan-2-ol 178

5-(4-morpholino-6-(pyrrolidin- 1-yl)pyrido[3,2-d]pyrimidin-2-yl)pyrimidin-2-amine 179

4-(1-((2-(1H- benzo[d]imidazol-4-yl)-4- morpholinopyrido[3,2-d]pyrimidin-6- yl)methyl)azetidin-3- yl)morpholine 180

4-(1-((2-(2-methyl-1H- benzo[d]imidazol-4-yl)-4- mropholinopyrido[3,2-d]pyrimidin-6- yl)methyl)azetidin-3- yl)morpholine 181

5-(6-isopropoxy-4- morpholinopyrido[3,2- d]pyrimidin-2-yl)pyrimidin-2-amine 182

5-(6-(azetidin-1-yl)-4- morpholinopyrido[3,2-d]pyrimidin-2-yl)pyrimidin-2- amine 183

5-(6-(3-methoxyazetidin-1-yl)- 4-morpholinopyrido[3,2-d]pyrimidin-2-yl)pyrimidin-2- amine 184

5-(6-(cyclobutylmethoxy)-4- morpholinopyrido[3,2-d]pyrimidin-2-yl)pyrimidin-2- amine 185

5-(6-(3-fluoroazetidin-1-yl)-4- morpholinopyrido[3,2-d]pyrimidin-2-yl)pyrimidin-2- amine 186

2-(2-aminopyrimidin-5-yl)- N,N-dimethyl-4- morpholinopyrido[3,2-d]pyrimidin-6-amine 187

5-(4,6- dimorpholinopyrido[3,2- d]pyrimidin-2-yl)pyrimidin-2- amine 188

5-(6-((3-methyloxetan-3- yl)methoxy)-4- morpholinopyrido[3,2-d]pyrimidin-2-yl)pyrimidin-2- amine 189

2-(2-aminopyrimidin-5-yl)- N,N-diethyl-4- morpholinopyrido[3,2-d]pyrimidin-6-amine 190

5-(6-(cyclopropylmethoxy)-4- morpholinopyrido[3,2-d]pyrimidin-2-yl)pyrimidin-2- amine 191

5-(6-(isopropylthio)-4- morpholinopyrido[3,2-d]pyrimidin-2-yl)pyrimidin-2- amine 192

5-(6-(2-methoxyethoxy)-4- morpholinopyrido[3,2-d]pyrimidin-2-yl)pyrimidin-2- amine 193

5-(4-morpholino-6-(2,2,2- trifluoroethoxy)pyrido[3,2-d]pyrimidin-2-yl)pyrimidin-2- amine 194

2-(2-aminopyrimidin-5-yl)-N- isopropyl-N-methyl-4- morpholinopyrido[3,2-d]pyrimidin-6-amine 195

5-(4-morpholine-6-(2-oxa-6- azaspiro[3.3]heptan-6-yl)pyrido[3,2-d]pyrimidin-2- yl)pyrimidin-2-amine 196

5-(6-cyclopropyl-4- morpholinopyrido[3,2- d]pyrimidin-2-yl)pyrimidin-2-amine 197

5-(7-methyl-4- morpholinopyrido[3,2- d]pyrimidin-2-yl)pyrimidin-2- amine198

5-(7-cyclopropyl-4- morpholinopyrido[3,2- d]pyrimidin-2-yl)pyrimidin-2-amine 199

4-(2,7-bis(3-methoxyazetidin- 1-yl)pyrido[3,2-d]pyrimidin-4-yl)morpholine 200

5-(7-(3-methoxyazetidin-1-yl)- 4-morpholinopyrido[3,2-d]pyrimidin-2-yl)pyrimidin-2- amine 201

5-(7-isopropoxy-4- morpholinopyrido[3,2- d]pyrimidin-2-yl)pyrimidin-2-amine

Administration of Formula I Compounds

The Formula I compounds of the invention may be administered by a routeappropriate to the condition to be treated. Suitable routes includeoral, parenteral (including subcutaneous, intramuscular, intravenous,intraarterial, intradermal, intrathecal and epidural), transdermal,rectal, nasal, topical (including buccal and sublingual), vaginal,intraperitoneal, intrapulmonary and intranasal. For localimmunosuppressive treatment, the compounds may be administered byintralesional administration, including perfusing or otherwisecontacting the graft with the inhibitor before transplantation. It willbe appreciated that the preferred route may vary with for example thecondition of the recipient. Where the compound is administered orally,it may be formulated as a pill, capsule, tablet, etc. with apharmaceutically acceptable carrier or excipient. Where the compound isadministered parenterally, it may be formulated with a pharmaceuticallyacceptable parenteral vehicle and in a unit dosage injectable form, asdetailed below.

A dose to treat human patients may range from about 10 mg to about 1000mg of Formula I compound. A typical dose may be about 100 mg to about300 mg of the compound. A dose may be administered once a day (QID),twice per day (BID), or more frequently, depending on thepharmacokinetic and pharmacodynamic properties, including absorption,distribution, metabolism, and excretion of the particular compound. Inaddition, toxicity factors may influence the dosage and administrationregimen. When administered orally, the pill, capsule, or tablet may beingested daily or less frequently for a specified period of time. Theregimen may be repeated for a number of cycles of therapy.

Methods of Treatment with Formula I Compounds

Formula I compounds of the present invention are useful for treating ahuman or animal patient suffering from a disease or disorder arisingfrom function or behavior associated with PI3 kinase, in particular withthe p110δ (delta) isoform of PI3 kinase such as an immune disorder,cardiovascular disease, viral infection, inflammation, ametabolism/endocrine disorder or a neurological disorder, may thus betreated by a method comprising the administration thereto of a compoundof the present invention as defined above. A human or animal patientsuffering from abnormal cell growth or cellular proliferative diseasessuch as tumor and/or cancerous cell growth mediated by PI3K may also betreated by a method comprising the administration thereto of a Formula Icompound. The condition of the patient may thereby be improved orameliorated. In particular, the compounds are useful in the treatment ofhuman or animal (e.g., murine) cancers, including, for example, breast,ovary, cervix, prostate, testis, genitourinary tract, esophagus, larynx,glioblastoma, neuroblastoma, stomach, skin, keratoacanthoma, lung,epidermoid carcinoma, large cell carcinoma, non-small cell lungcarcinoma (NSCLC), small cell carcinoma, lung adenocarcinoma, bone,colon, adenoma, pancreas, adenocarcinoma, thyroid, follicular carcinoma,undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma,sarcoma, bladder carcinoma, liver carcinoma and biliary passages, kidneycarcinoma, pancreatic, myeloid disorders, lymphoma, hairy cells, buccalcavity, naso-pharyngeal, pharynx, lip, tongue, mouth, small intestine,colon-rectum, large intestine, rectum, brain and central nervous system,Hodgkin's, leukemia, bronchus, thyroid, liver and intrahepatic bileduct, hepatocellular, gastric, glioma/glioblastoma, endometrial,melanoma, kidney and renal pelvis, urinary bladder, uterine corpus,uterine cervix, multiple myeloma, acute myelogenous leukemia, chronicmyelogenous leukemia, lymphocytic leukemia, myeloid leukemia, oralcavity and pharynx, non-Hodgkin lymphoma, melanoma, and villous colonadenoma.

Formula I compounds may be useful for in vitro, in situ, and in vivodiagnosis or treatment of mammalian cells, organisms, or associatedpathological conditions, such as systemic and local inflammation,immune-inflammatory diseases such as rheumatoid arthritis, immunesuppression, organ transplant rejection, allergies, ulcerative colitis,Crohn's disease, dermatitis, asthma, systemic lupus erythematosus,Sjögren's Syndrome, multiple sclerosis, scleroderma/systemic sclerosis,idiopathic thrombocytopenic purpura (ITP), anti-neutrophil cytoplasmicantibodies (ANCA) vasculitis, chronic obstructive pulmonary disease(COPD), psoriasis, and for general joint protective effects.

Methods of the invention also include treating such diseases asarthritic diseases, such as rheumatoid arthritis, monoarticulararthritis, osteoarthritis, gouty arthritis, spondylitis; Behcet disease;sepsis, septic shock, endotoxic shock, gram negative sepsis, grampositive sepsis, and toxic shock syndrome; multiple organ injurysyndrome secondary to septicemia, trauma, or hemorrhage; ophthalmicdisorders such as allergic conjunctivitis, vernal conjunctivitis,uveitis, and thyroid-associated ophthalmopathy; eosinophilic granuloma;pulmonary or respiratory disorders such as asthma, chronic bronchitis,allergic rhinitis, ARDS, chronic pulmonary inflammatory disease (e.g.,chronic obstructive pulmonary disease), silicosis, pulmonarysarcoidosis, pleurisy, alveolitis, vasculitis, emphysema, pneumonia,bronchiectasis, and pulmonary oxygen toxicity; reperfusion injury of themyocardium, brain, or extremities; fibrosis such as cystic fibrosis;keloid formation or scar tissue formation; atherosclerosis; autoimmunediseases, such as systemic lupus erythematosus (SLE), autoimmunethyroiditis, multiple sclerosis, some forms of diabetes, and Reynaud'ssyndrome; and transplant rejection disorders such as GVHD and allograftrejection; chronic glomerulonephritis; inflammatory bowel diseases suchas chronic inflammatory bowel disease (CIBD), Crohn's disease,ulcerative colitis, and necrotizing enterocolitis; inflammatorydermatoses such as contact dermatitis, atopic dermatitis, psoriasis, orurticaria; fever and myalgias due to infection; central or peripheralnervous system inflammatory disorders such as meningitis, encephalitis,and brain or spinal cord injury due to minor trauma; Sjögren's syndrome;diseases involving leukocyte diapedesis; alcoholic hepatitis; bacterialpneumonia; antigen-antibody complex mediated diseases; hypovolemicshock; Type I diabetes mellitus; acute and delayed hypersensitivity;disease states due to leukocyte dyscrasia and metastasis; thermalinjury; granulocyte transfusion-associated syndromes; andcytokine-induced toxicity.

The methods of the invention can have utility in treating subjects whoare or can be subject to reperfusion injury, i.e., injury resulting fromsituations in which a tissue or organ experiences a period of ischemiafollowed by reperfusion. The term “ischemia” refers to localized tissueanemia due to obstruction of the inflow of arterial blood. Transientischemia followed by reperfusion characteristically results inneutrophil activation and transmigration through the endothelium of theblood vessels in the affected area. Accumulation of activatedneutrophils in turn results in generation of reactive oxygenmetabolites, which damage components of the involved tissue or organ.This phenomenon of “reperfusion injury” is commonly associated withconditions such as vascular stroke (including global and focalischemia), hemorrhagic shock, myocardial ischemia or infarction, organtransplantation, and cerebral vasospasm. To illustrate, reperfusioninjury occurs at the termination of cardiac bypass procedures or duringcardiac arrest when the heart, once prevented from receiving blood,begins to reperfuse. It is expected that inhibition of PI3K deltaactivity may result in reduced amounts of reperfusion injury in suchsituations.

Methods of the invention also include treating cancer in a mammalcomprised of administering to said mammal a therapeutically effectiveamount of a Formula I compound wherein the cancer is breast, ovary,cervix, prostate, testis, genitourinary tract, esophagus, larynx,glioblastoma, neuroblastoma, stomach, skin, keratoacanthoma, lung,epidermoid carcinoma, large cell carcinoma, non-small cell lungcarcinoma (NSCLC), small cell carcinoma, lung adenocarcinoma, bone,colon, adenoma, pancreas, adenocarcinoma, thyroid, follicular carcinoma,undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma,sarcoma, bladder carcinoma, liver carcinoma and biliary passages, kidneycarcinoma, pancreatic, myeloid disorders, lymphoma, hairy cells, buccalcavity, naso-pharyngeal, pharynx, lip, tongue, mouth, small intestine,colon-rectum, large intestine, rectum, brain and central nervous system,Hodgkin's or leukemia.

Formula I compounds may be useful for treating conditions of the brainand central nervous system which require transport across theblood-brain barrier. Certain Formula I compounds have favorablepenetrant properties across the blood-brain barrier for delivery to thebrain. Disorders of the brain which may be effectively treated withFormula I compounds include metastatic and primary brain tumors, such asglioblastoma and melanoma.

Pharmaceutical Formulations

In order to use a Formula I compound for the therapeutic treatment(including prophylactic treatment) of mammals including humans, it isnormally formulated in accordance with standard pharmaceutical practiceas a pharmaceutical composition. According to this aspect of theinvention there is provided a pharmaceutical composition comprising acompound of this invention in association with a pharmaceuticallyacceptable diluent or carrier.

A typical formulation is prepared by mixing a compound of the presentinvention and a carrier, diluent or excipient. Suitable carriers,diluents and excipients are well known to those skilled in the art andinclude materials such as carbohydrates, waxes, water soluble and/orswellable polymers, hydrophilic or hydrophobic materials, gelatin, oils,solvents, water and the like. The particular carrier, diluent orexcipient used will depend upon the means and purpose for which thecompound of the present invention is being applied. Solvents aregenerally selected based on solvents recognized by persons skilled inthe art as safe (GRAS) to be administered to a mammal. In general, safesolvents are non-toxic aqueous solvents such as water and othernon-toxic solvents that are soluble or miscible in water. Suitableaqueous solvents include water, ethanol, propylene glycol, polyethyleneglycols (e.g., PEG 400, PEG 300), etc. and mixtures thereof. Theformulations may also include one or more buffers, stabilizing agents,surfactants, wetting agents, lubricating agents, emulsifiers, suspendingagents, preservatives, antioxidants, opaquing agents, glidants,processing aids, colorants, sweeteners, perfuming agents, flavoringagents and other known additives to provide an elegant presentation ofthe drug (i.e., a compound of the present invention or pharmaceuticalcomposition thereof) or aid in the manufacturing of the pharmaceuticalproduct (i.e., medicament).

The choice of formulation depends on various factors such as the mode ofdrug administration and bioavailability of the drug substance. Fordelivery via inhalation the compound can be formulated as liquidsolution, suspensions, aerosol propellants or dry powder and loaded intoa suitable dispenser for administration. There are several types ofpharmaceutical inhalation devices-nebulizer inhalers, metered doseinhalers (MDI) and dry powder inhalers (DPI). Nebulizer devices producea stream of high velocity air that causes the therapeutic agents (whichare formulated in a liquid form) to spray as a mist that is carried intothe patient's respiratory tract. MDI's typically are formulationpackaged with a compressed gas. Upon actuation, the device discharges ameasured amount of therapeutic agent by compressed gas, thus affording areliable method of administering a set amount of agent. DPI dispensestherapeutic agents in the form of a free flowing powder that can bedispersed in the patient's inspiratory air-stream during breathing bythe device. In order to achieve a free flowing powder, the therapeuticagent is formulated with an excipient such as lactose. A measured amountof the therapeutic agent is stored in a capsule form and is dispensedwith each actuation.

Formulations include where the particle size of a Formula I compound isbetween 10-1000 nm, or between 10-400 nm. Such pharmaceuticalformulations may be useful for a Formula I compound with poorbioavailability based upon the principle that bioavailability can beincreased by increasing the surface area by decreasing particle size(U.S. Pat. No. 4,107,288, U.S. Pat. No. 5,145,684) to the size rangefrom 10 to 1,000 nm where the Formula I compound is supported on a crosslinked matrix of macromolecules.

The formulations may be prepared using conventional dissolution andmixing procedures. For example, the bulk drug substance (i.e., compoundof the present invention or stabilized form of the compound (e.g.,complex with a cyclodextrin derivative or other known complexationagent) is dissolved in a suitable solvent in the presence of one or moreof the excipients described above. The compound of the present inventionis typically formulated into pharmaceutical dosage forms to provide aneasily controllable dosage of the drug and to enable patient compliancewith the prescribed regimen.

The pharmaceutical composition (or formulation) for application may bepackaged in a variety of ways depending upon the method used foradministering the drug. Generally, an article for distribution includesa container having deposited therein the pharmaceutical formulation inan appropriate form. Suitable containers are well known to those skilledin the art and include materials such as bottles (plastic and glass),sachets, ampoules, plastic bags, metal cylinders, and the like. Thecontainer may also include a tamper-proof assemblage to preventindiscreet access to the contents of the package. In addition, thecontainer has deposited thereon a label that describes the contents ofthe container. The label may also include appropriate warnings.

Pharmaceutical formulations of the compounds of the present inventionmay be prepared for various routes and types of administration. Forexample, a compound of Formula I having the desired degree of purity mayoptionally be mixed with pharmaceutically acceptable diluents, carriers,excipients or stabilizers (Remington's Pharmaceutical Sciences (1980)16^(th) edition, Osol, A. Ed.), in the form of a lyophilizedformulation, milled powder, or an aqueous solution. Formulation may beconducted by mixing at ambient temperature at the appropriate pH, and atthe desired degree of purity, with physiologically acceptable carriers,i.e., carriers that are non-toxic to recipients at the dosages andconcentrations employed. The pH of the formulation depends mainly on theparticular use and the concentration of compound, but may range fromabout 3 to about 8. Formulation in an acetate buffer at pH 5 is asuitable embodiment.

The compound ordinarily can be stored as a solid composition, alyophilized formulation or as an aqueous solution.

The pharmaceutical compositions of the invention will be formulated,dosed and administered in a fashion, i.e., amounts, concentrations,schedules, course, vehicles and route of administration, consistent withgood medical practice. Factors for consideration in this context includethe particular disorder being treated, the particular mammal beingtreated, the clinical condition of the individual patient, the cause ofthe disorder, the site of delivery of the agent, the method ofadministration, the scheduling of administration, and other factorsknown to medical practitioners. The “therapeutically effective amount”of the compound to be administered will be governed by suchconsiderations, and is the minimum amount necessary to prevent,ameliorate, or treat the hyperproliferative disorder.

As a general proposition, the initial pharmaceutically effective amountof the inhibitor administered parenterally per dose will be in the rangeof about 0.01-100 mg/kg, namely about 0.1 to 20 mg/kg of patient bodyweight per day, with the typical initial range of compound used being0.3 to 15 mg/kg/day.

Acceptable diluents, carriers, excipients and stabilizers are nontoxicto recipients at the dosages and concentrations employed, and includebuffers such as phosphate, citrate and other organic acids; antioxidantsincluding ascorbic acid and methionine; preservatives (such asoctadecyldimethylbenzyl ammonium chloride; hexamethonium chloride;benzalkonium chloride, benzethonium chloride; phenol, butyl or benzylalcohol; alkyl parabens such as methyl or propyl paraben; catechol;resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecularweight (less than about 10 residues) polypeptides; proteins, such asserum albumin, gelatin, or immunoglobulins; hydrophilic polymers such aspolyvinylpyrrolidone; amino acids such as glycine, glutamine,asparagine, histidine, arginine, or lysine; monosaccharides,disaccharides and other carbohydrates including glucose, mannose, ordextrins; chelating agents such as EDTA; sugars such as sucrose,mannitol, trehalose or sorbitol; salt-forming counter-ions such assodium; metal complexes (e.g., Zn-protein complexes); and/or non-ionicsurfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG). Theactive pharmaceutical ingredients may also be entrapped in microcapsulesprepared, for example, by coacervation techniques or by interfacialpolymerization, for example, hydroxymethylcellulose orgelatin-microcapsules and poly-(methylmethacylate) microcapsules,respectively, in colloidal drug delivery systems (for example,liposomes, albumin microspheres, microemulsions, nano-particles andnanocapsules) or in macroemulsions. Such techniques are disclosed inRemington's Pharmaceutical Sciences 16^(th) edition, Osol, A. Ed.(1980).

Sustained-release preparations of compounds of Formula I may beprepared. Suitable examples of sustained-release preparations includesemipermeable matrices of solid hydrophobic polymers containing acompound of Formula I, which matrices are in the form of shapedarticles, e.g., films, or microcapsules. Examples of sustained-releasematrices include polyesters, hydrogels (for example,poly(2-hydroxyethyl-methacrylate), or poly(vinyl alcohol)), polylactides(U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid andgamma-ethyl-L-glutamate, non-degradable ethylene-vinyl acetate,degradable lactic acid-glycolic acid copolymers such as the LUPRONDEPOT™ (injectable microspheres composed of lactic acid-glycolic acidcopolymer and leuprolide acetate) and poly-D-(−)-3-hydroxybutyric acid.

The formulations include those suitable for the administration routesdetailed herein. The formulations may conveniently be presented in unitdosage form and may be prepared by any of the methods well known in theart of pharmacy. Techniques and formulations generally are found inRemington's Pharmaceutical Sciences (Mack Publishing Co., Easton, Pa.).Such methods include the step of bringing into association the activeingredient with the carrier which constitutes one or more accessoryingredients. In general the formulations are prepared by uniformly andintimately bringing into association the active ingredient with liquidcarriers or finely divided solid carriers or both, and then, ifnecessary, shaping the product.

Formulations of a compound of Formula I suitable for oral administrationmay be prepared as discrete units such as pills, capsules, cachets ortablets each containing a predetermined amount of a compound of FormulaI. Compressed tablets may be prepared by compressing in a suitablemachine the active ingredient in a free-flowing form such as a powder orgranules, optionally mixed with a binder, lubricant, inert diluent,preservative, surface active or dispersing agent. Molded tablets may bemade by molding in a suitable machine a mixture of the powdered activeingredient moistened with an inert liquid diluent. The tablets mayoptionally be coated or scored and optionally are formulated so as toprovide slow or controlled release of the active ingredient therefrom.Tablets, troches, lozenges, aqueous or oil suspensions, dispersiblepowders or granules, emulsions, hard or soft capsules, e.g., gelatincapsules, syrups or elixirs may be prepared for oral use. Formulationsof compounds of Formula I intended for oral use may be preparedaccording to any method known to the art for the manufacture ofpharmaceutical compositions and such compositions may contain one ormore agents including sweetening agents, flavoring agents, coloringagents and preserving agents, in order to provide a palatablepreparation. Tablets containing the active ingredient in admixture withnon-toxic pharmaceutically acceptable excipient which are suitable formanufacture of tablets are acceptable. These excipients may be, forexample, inert diluents, such as calcium or sodium carbonate, lactose,calcium or sodium phosphate; granulating and disintegrating agents, suchas maize starch, or alginic acid; binding agents, such as starch,gelatin or acacia; and lubricating agents, such as magnesium stearate,stearic acid or talc. Tablets may be uncoated or may be coated by knowntechniques including microencapsulation to delay disintegration andadsorption in the gastrointestinal tract and thereby provide a sustainedaction over a longer period. For example, a time delay material such asglyceryl monostearate or glyceryl distearate alone or with a wax may beemployed.

For treatment of the eye or other external tissues, e.g., mouth andskin, the formulations are preferably applied as a topical ointment orcream containing the active ingredient(s) in an amount of, for example,0.075 to 20% w/w. When formulated in an ointment, the active ingredientsmay be employed with either a paraffinic or a water-miscible ointmentbase. Alternatively, the active ingredients may be formulated in a creamwith an oil-in-water cream base. If desired, the aqueous phase of thecream base may include a polyhydric alcohol, i.e., an alcohol having twoor more hydroxyl groups such as propylene glycol, butane 1,3-diol,mannitol, sorbitol, glycerol and polyethylene glycol (including PEG 400)and mixtures thereof. The topical formulations may desirably include acompound which enhances absorption or penetration of the activeingredient through the skin or other affected areas. Examples of suchdermal penetration enhancers include dimethyl sulfoxide and relatedanalogs. The oily phase of the emulsions of this invention may beconstituted from known ingredients in a known manner, including amixture of at least one emulsifier with a fat or an oil, or with both afat and an oil. A hydrophilic emulsifier included together with alipophilic emulsifier acts as a stabilizer. Together, the emulsifier(s)with or without stabilizer(s) make up the so-called emulsifying wax, andthe wax together with the oil and fat make up the so-called emulsifyingointment base which forms the oily dispersed phase of the creamformulations. Emulsifiers and emulsion stabilizers suitable for use inthe formulation of the invention include Tween® 60, Span® 80,cetostearyl alcohol, benzyl alcohol, myristyl alcohol, glycerylmono-stearate and sodium lauryl sulfate.

Aqueous suspensions of Formula I compounds contain the active materialsin admixture with excipients suitable for the manufacture of aqueoussuspensions. Such excipients include a suspending agent, such as sodiumcarboxymethylcellulose, croscarmellose, povidone, methylcellulose,hydroxypropyl methylcellulose, sodium alginate, polyvinylpyrrolidone,gum tragacanth and gum acacia, and dispersing or wetting agents such asa naturally occurring phosphatide (e.g., lecithin), a condensationproduct of an alkylene oxide with a fatty acid (e.g., polyoxyethylenestearate), a condensation product of ethylene oxide with a long chainaliphatic alcohol (e.g., heptadecaethyleneoxycetanol), a condensationproduct of ethylene oxide with a partial ester derived from a fatty acidand a hexitol anhydride (e.g., polyoxyethylene sorbitan monooleate). Theaqueous suspension may also contain one or more preservatives such asethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one ormore flavoring agents and one or more sweetening agents, such as sucroseor saccharin.

The pharmaceutical compositions of compounds of Formula I may be in theform of a sterile injectable preparation, such as a sterile injectableaqueous or oleaginous suspension. This suspension may be formulatedaccording to the known art using those suitable dispersing or wettingagents and suspending agents which have been mentioned above. Thesterile injectable preparation may also be a sterile injectable solutionor suspension in a non-toxic parenterally acceptable diluent or solvent,such as a solution in 1,3-butanediol or prepared as a lyophilizedpowder. Among the acceptable vehicles and solvents that may be employedare water, Ringer's solution and isotonic sodium chloride solution. Inaddition, sterile fixed oils may conventionally be employed as a solventor suspending medium. For this purpose any bland fixed oil may beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid may likewise be used in the preparation ofinjectables.

The amount of active ingredient that may be combined with the carriermaterial to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. For example, atime-release formulation intended for oral administration to humans maycontain approximately 1 to 1000 mg of active material compounded with anappropriate and convenient amount of carrier material which may varyfrom about 5 to about 95% of the total compositions (weight:weight). Thepharmaceutical composition can be prepared to provide easily measurableamounts for administration. For example, an aqueous solution intendedfor intravenous infusion may contain from about 3 to 500 μg of theactive ingredient per milliliter of solution in order that infusion of asuitable volume at a rate of about 30 mL/hr (hour) can occur.

Formulations suitable for parenteral administration include aqueous andnon-aqueous sterile injection solutions which may contain anti-oxidants,buffers, bacteriostats and solutes which render the formulation isotonicwith the blood of the intended recipient; and aqueous and non-aqueoussterile suspensions which may include suspending agents and thickeningagents.

Formulations suitable for topical administration to the eye also includeeye drops wherein the active ingredient is dissolved or suspended in asuitable carrier, especially an aqueous solvent for the activeingredient. The active ingredient is preferably present in suchformulations in a concentration of about 0.5 to 20% w/w, for exampleabout 0.5 to 10% w/w, for example about 1.5% w/w.

Formulations suitable for topical administration in the mouth includelozenges comprising the active ingredient in a flavored basis, usuallysucrose and acacia or tragacanth; pastilles comprising the activeingredient in an inert basis such as gelatin and glycerin, or sucroseand acacia; and mouthwashes comprising the active ingredient in asuitable liquid carrier.

Formulations for rectal administration may be presented as a suppositorywith a suitable base comprising for example cocoa butter or asalicylate.

Formulations suitable for intrapulmonary or nasal administration have aparticle size for example in the range of 0.1 to 500 microns (includingparticle sizes in a range between 0.1 and 500 microns in incrementsmicrons such as 0.5, 1, 30 microns, 35 microns, etc.), which isadministered by rapid inhalation through the nasal passage or byinhalation through the mouth so as to reach the alveolar sacs. Suitableformulations include aqueous or oily solutions of the active ingredient.Formulations suitable for aerosol or dry powder administration may beprepared according to conventional methods and may be delivered withother therapeutic agents such as compounds heretofore used in thetreatment or prophylaxis disorders as described below.

Formulations suitable for vaginal administration may be presented aspessaries, tampons, creams, gels, pastes, foams or spray formulationscontaining in addition to the active ingredient such carriers as areknown in the art to be appropriate.

The formulations may be packaged in unit-dose or multi-dose containers,for example sealed ampoules and vials, and may be stored in afreeze-dried (lyophilized) condition requiring only the addition of thesterile liquid carrier, for example water, for injection immediatelyprior to use. Extemporaneous injection solutions and suspensions areprepared from sterile powders, granules and tablets of the kindpreviously described. Preferred unit dosage formulations are thosecontaining a daily dose or unit daily sub-dose, as herein above recited,or an appropriate fraction thereof, of the active ingredient.

The invention further provides veterinary compositions comprising atleast one active ingredient as above defined together with a veterinarycarrier therefore. Veterinary carriers are materials useful for thepurpose of administering the composition and may be solid, liquid orgaseous materials which are otherwise inert or acceptable in theveterinary art and are compatible with the active ingredient. Theseveterinary compositions may be administered parenterally, orally or byany other desired route.

Combination Therapy

The compounds of Formula I may be employed alone or in combination withother therapeutic agents for the treatment of a disease or disorderdescribed herein, such as inflammation or a hyperproliferative disorder(e.g., cancer). In certain embodiments, a compound of Formula I iscombined in a pharmaceutical combination formulation, or dosing regimenas combination therapy, with a second therapeutic compound that hasanti-inflammatory or anti-hyperproliferative properties or that isuseful for treating an inflammation, immune-response disorder, orhyperproliferative disorder (e.g., cancer). The second therapeutic agentmay be an NSAID anti-inflammatory agent. The second therapeutic agentmay be a chemotherapeutic agent. The second compound of thepharmaceutical combination formulation or dosing regimen preferably hascomplementary activities to the compound of Formula I such that they donot adversely affect each other. Such compounds are suitably present incombination in amounts that are effective for the purpose intended. Inone embodiment, a composition of this invention comprises a compound ofFormula I, or a stereoisomer, geometric isomer, tautomer, solvate,metabolite, or pharmaceutically acceptable salt or prodrug thereof, incombination with a therapeutic agent such as an NSAID.

Combination refers to either a fixed combination in one dosage unitform, or a kit of parts for the combined administration where a compoundof Formula I and a second therapeutic agent may be administeredindependently at the same time or separately within time intervals,especially where these time intervals allow that the combinationpartners show a cooperative, e.g. synergistic effect. The combinationtherapy methods and compositions of the invention are meant to encompassadministration of the selected combination to a single subject in needthereof (e.g. a patient), and are intended to include treatment regimensin which the agents are not necessarily administered by the same routeof administration or at the same time. The term “pharmaceuticalcombination” means a product that results from the mixing or combiningof more than one active ingredient and includes both fixed and non-fixedcombinations of the active ingredients. The term “fixed combination”means that the active ingredients, e.g. a compound of Formula I and asecond therapeutic combination partner, are both administered to apatient simultaneously in the form of a single entity or dosage. Theterm “non-fixed combination” means that the active ingredients, e.g. acompound of Formula I and a second therapeutic combination partner, areboth administered to a patient as separate entities eithersimultaneously, concurrently or sequentially with no specific timelimits, wherein such administration provides therapeutically effectivelevels of the two compounds in the body of the patient. The latter alsoapplies to cocktail therapy, e.g. the administration of three or moreactive ingredients.

The combination therapy may be administered as a simultaneous orsequential regimen. When administered sequentially, the combination maybe administered in two or more administrations. The combinedadministration includes coadministration, using separate formulations ora single pharmaceutical formulation, and consecutive administration ineither order, wherein preferably there is a time period while both (orall) active agents simultaneously exert their biological activities.

Suitable dosages for any of the above coadministered agents are thosepresently used and may be lowered due to the combined action (synergy)of the newly identified agent and other therapeutic agents ortreatments.

The combination therapy may provide “synergy” and prove “synergistic”,i.e., the effect achieved when the active ingredients used together isgreater than the sum of the effects that results from using thecompounds separately. A synergistic effect may be attained when theactive ingredients are: (1) co-formulated and administered or deliveredsimultaneously in a combined, unit dosage formulation; (2) delivered byalternation or in parallel as separate formulations; or (3) by someother regimen. When delivered in alternation therapy, a synergisticeffect may be attained when the compounds are administered or deliveredsequentially, e.g., by different injections in separate syringes,separate pills or capsules, or separate infusions. In general, duringalternation therapy, an effective dosage of each active ingredient isadministered sequentially, i.e., serially, whereas in combinationtherapy, effective dosages of two or more active ingredients areadministered together.

In a particular embodiment of therapy, a compound of Formula I, or astereoisomer, geometric isomer, tautomer, solvate, metabolite, orpharmaceutically acceptable salt or prodrug thereof, may be combinedwith other therapeutic, hormonal or antibody agents such as thosedescribed herein, as well as combined with surgical therapy andradiotherapy. Combination therapies according to the present inventionthus comprise the administration of at least one compound of Formula I,or a stereoisomer, geometric isomer, tautomer, solvate, metabolite, orpharmaceutically acceptable salt or prodrug thereof, and the use of atleast one other cancer treatment method. The amounts of the compound(s)of Formula I and the other pharmaceutically active chemotherapeuticagent(s) and the relative timings of administration will be selected inorder to achieve the desired combined therapeutic effect.

Metabolites of Compounds of Formula I

Also falling within the scope of this invention are the in vivometabolic products of Formula I described herein. Such products mayresult for example from the oxidation, reduction, hydrolysis, amidation,deamidation, esterification, deesterification, enzymatic cleavage, andthe like, of the administered compound. Accordingly, the inventionincludes metabolites of compounds of Formula I, including compoundsproduced by a process comprising contacting a compound of this inventionwith a mammal for a period of time sufficient to yield a metabolicproduct thereof.

Metabolite products typically are identified by preparing aradiolabelled (e.g., ¹⁴C or ³H) isotope of a compound of the invention,administering it parenterally in a detectable dose (e.g., greater thanabout 0.5 mg/kg) to an animal such as rat, mouse, guinea pig, monkey, orto man, allowing sufficient time for metabolism to occur (typicallyabout 30 seconds to 30 hours) and isolating its conversion products fromthe urine, blood or other biological samples. These products are easilyisolated since they are labeled (others are isolated by the use ofantibodies capable of binding epitopes surviving in the metabolite). Themetabolite structures are determined in conventional fashion, e.g., byMS, LC/MS or NMR analysis. In general, analysis of metabolites is donein the same way as conventional drug metabolism studies well known tothose skilled in the art. The metabolite products, so long as they arenot otherwise found in vivo, are useful in diagnostic assays fortherapeutic dosing of the compounds of the invention.

Articles of Manufacture

In another embodiment of the invention, an article of manufacture, or“kit”, containing materials useful for the treatment of the diseases anddisorders described above is provided. In one embodiment, the kitcomprises a container comprising a compound of Formula I. The kit mayfurther comprise a label or package insert, on or associated with thecontainer. The term “package insert” is used to refer to instructionscustomarily included in commercial packages of therapeutic products,that contain information about the indications, usage, dosage,administration, contraindications and/or warnings concerning the use ofsuch therapeutic products. Suitable containers include, for example,bottles, vials, syringes, blister pack, etc. The container may be formedfrom a variety of materials such as glass or plastic. The container mayhold a compound of Formula I or a formulation thereof which is effectivefor treating the condition and may have a sterile access port (forexample, the container may be an intravenous solution bag or a vialhaving a stopper pierceable by a hypodermic injection needle). At leastone active agent in the composition is a compound of Formula I. Thelabel or package insert indicates that the composition is used fortreating the condition of choice, such as cancer. In addition, the labelor package insert may indicate that the patient to be treated is onehaving a disorder such as a hyperproliferative disorder,neurodegeneration, cardiac hypertrophy, pain, migraine or aneurotraumatic disease or event. In one embodiment, the label or packageinserts indicates that the composition comprising a compound of FormulaI can be used to treat a disorder resulting from abnormal cell growth.The label or package insert may also indicate that the composition canbe used to treat other disorders. Alternatively, or additionally, thearticle of manufacture may further comprise a second containercomprising a pharmaceutically acceptable buffer, such as bacteriostaticwater for injection (BWFI), phosphate-buffered saline, Ringer's solutionand dextrose solution. It may further include other materials desirablefrom a commercial and user standpoint, including other buffers,diluents, filters, needles, and syringes.

The kit may further comprise directions for the administration of thecompound of Formula I and, if present, the second pharmaceuticalformulation. For example, if the kit comprises a first compositioncomprising a compound of Formula I and a second pharmaceuticalformulation, the kit may further comprise directions for thesimultaneous, sequential or separate administration of the first andsecond pharmaceutical compositions to a patient in need thereof.

In another embodiment, the kits are suitable for the delivery of solidoral forms of a compound of Formula I, such as tablets or capsules. Sucha kit preferably includes a number of unit dosages. Such kits caninclude a card having the dosages oriented in the order of theirintended use. An example of such a kit is a “blister pack”. Blisterpacks are well known in the packaging industry and are widely used forpackaging pharmaceutical unit dosage forms. If desired, a memory aid canbe provided, for example in the form of numbers, letters, or othermarkings or with a calendar insert, designating the days in thetreatment schedule in which the dosages can be administered.

According to one embodiment, a kit may comprise (a) a first containerwith a compound of Formula I contained therein; and optionally (b) asecond container with a second pharmaceutical formulation containedtherein, wherein the second pharmaceutical formulation comprises asecond compound with anti-hyperproliferative activity. Alternatively, oradditionally, the kit may further comprise a third container comprisinga pharmaceutically-acceptable buffer, such as bacteriostatic water forinjection (BWFI), phosphate-buffered saline, Ringer's solution anddextrose solution. It may further include other materials desirable froma commercial and user standpoint, including other buffers, diluents,filters, needles, and syringes.

In certain other embodiments wherein the kit comprises a composition ofFormula I and a second therapeutic agent, the kit may comprise acontainer for containing the separate compositions such as a dividedbottle or a divided foil packet, however, the separate compositions mayalso be contained within a single, undivided container. Typically, thekit comprises directions for the administration of the separatecomponents. The kit form is particularly advantageous when the separatecomponents are preferably administered in different dosage forms (e.g.,oral and parenteral), are administered at different dosage intervals, orwhen titration of the individual components of the combination isdesired by the prescribing physician.

Preparation of Formula I Compounds

Compounds of Formula I may be synthesized by synthetic routes thatinclude processes analogous to those well-known in the chemical arts,particularly in light of the description contained herein, and those forother heterocycles described in: Comprehensive Heterocyclic ChemistryII, Editors Katritzky and Rees, Elsevier, 1997, e.g. Volume 3; LiebigsAnnalen der Chemie, (9):1910-16, (1985); Helvetica Chimica Acta,41:1052-60, (1958); Arzneimittel-Forschung, 40(12):1328-31, (1990), eachof which are expressly incorporated by reference. Starting materials aregenerally available from commercial sources such as Aldrich Chemicals(Milwaukee, Wis.) or are readily prepared using methods well known tothose skilled in the art (e.g., prepared by methods generally describedin Louis F. Fieser and Mary Fieser, Reagents for Organic Synthesis, v.1-23, Wiley, N.Y. (1967-2006 ed.), or Beilsteins Handbuch derorganischen Chemie, 4, Aufl. ed. Springer-Verlag, Berlin, includingsupplements (also available via the Beilstein online database).

For certain embodiments, Formula I compounds may be readily preparedusing well-known procedures to prepare pyrido[3,2-d]pyrimidinecompounds, including the route of Scheme 1 and the methods of:Srinivasan and Broom (1981) J. Org. Chem. 46:1777-1781; Malagu K. et al(2009) Bioorg. & Med. Chem. Letters 19(20):5950-5953; Hayakawa M. et al(2006) Bioorg. & Med. Chem. 14(20):6847-6858; Nishikawa, K. et al (1976)Chem. and Pharm. Bull. 24(9):2057-2077; US 2008/0004285; US2009/0324543; US 2009/0131414; U.S. Pat. No. 6,608,053; and U.S. Pat.No. 3,939,268.

Synthetic chemistry transformations and protecting group methodologies(protection and deprotection) useful in synthesizing Formula I compoundsand necessary reagents and intermediates are known in the art andinclude, for example, those described in R. Larock, ComprehensiveOrganic Transformations, VCH Publishers (1989); T. W. Greene and P. G.M. Wuts, Protective Groups in Organic Synthesis, 3^(rd) Ed., John Wileyand Sons (1999); and L. Paquette, ed., Encyclopedia of Reagents forOrganic Synthesis, John Wiley and Sons (1995) and subsequent editionsthereof.

Compounds of Formula I may be prepared singly or as compound librariescomprising at least 2, for example 5 to 1,000 compounds, or 10 to 100compounds. Libraries of compounds of Formula I may be prepared by acombinatorial ‘split and mix’ approach or by multiple parallel synthesesusing either solution phase or solid phase chemistry, by proceduresknown to those skilled in the art. Thus according to a further aspect ofthe invention there is provided a compound library comprising at least 2compounds, or pharmaceutically acceptable salts thereof.

General Procedures and Examples provide exemplary methods for preparingFormula I compounds. Those skilled in the art will appreciate that othersynthetic routes may be used to synthesize the Formula I compounds.Although specific starting materials and reagents are depicted anddiscussed in the Schemes, General Procedures, and Examples, otherstarting materials and reagents can be easily substituted to provide avariety of derivatives and/or reaction conditions. In addition, many ofthe exemplary compounds prepared by the described methods can be furthermodified in light of this disclosure using conventional chemistry wellknown to those skilled in the art.

In preparing compounds of Formulas I, protection of remote functionality(e.g., primary or secondary amine) of intermediates may be necessary.The need for such protection will vary depending on the nature of theremote functionality and the conditions of the preparation methods.Suitable amino-protecting groups include acetyl, trifluoroacetyl,t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBz) and9-fluorenylmethyleneoxycarbonyl (Fmoc). The need for such protection isreadily determined by one skilled in the art. For a general descriptionof protecting groups and their use, see T. W. Greene, Protective Groupsin Organic Synthesis, John Wiley & Sons, New York, 1991.

Methods of Separation

In the methods of preparing Formula I compounds, it may be advantageousto separate reaction products from one another and/or from startingmaterials. The desired products of each step or series of steps isseparated and/or purified to the desired degree of homogeneity by thetechniques common in the art. Typically such separations involvemultiphase extraction, crystallization from a solvent or solventmixture, distillation, sublimation, or chromatography. Chromatographycan involve any number of methods including, for example: reverse-phaseand normal phase; size exclusion; ion exchange; high, medium and lowpressure liquid chromatography methods and apparatus; small scaleanalytical; simulated moving bed (SMB) and preparative thin or thicklayer chromatography, as well as techniques of small scale thin layerand flash chromatography.

Another class of separation methods involves treatment of a mixture witha reagent selected to bind to or render otherwise separable a desiredproduct, unreacted starting material, reaction by product, or the like.Such reagents include adsorbents or absorbents such as activated carbon,molecular sieves, ion exchange media, or the like. Alternatively, thereagents can be acids in the case of a basic material, bases in the caseof an acidic material, binding reagents such as antibodies, bindingproteins, selective chelators such as crown ethers, liquid/liquid ionextraction reagents (LIX), or the like. Selection of appropriate methodsof separation depends on the nature of the materials involved, such as,boiling point and molecular weight in distillation and sublimation,presence or absence of polar functional groups in chromatography,stability of materials in acidic and basic media in multiphaseextraction, and the like.

Diastereomeric mixtures can be separated into their individualdiastereomers on the basis of their physical chemical differences bymethods well known to those skilled in the art, such as bychromatography and/or fractional crystallization. Enantiomers can beseparated by converting the enantiomeric mixture into a diastereomericmixture by reaction with an appropriate optically active compound (e.g.,chiral auxiliary such as a chiral alcohol or Mosher's acid chloride),separating the diastereomers and converting (e.g., hydrolyzing) theindividual diastereoisomers to the corresponding pure enantiomers. Also,some of the compounds of the present invention may be atropisomers(e.g., substituted biaryls) and are considered as part of thisinvention. Enantiomers can also be separated by use of a chiral HPLCcolumn.

A single stereoisomer, e.g., an enantiomer, substantially free of itsstereoisomer may be obtained by resolution of the racemic mixture usinga method such as formation of diastereomers using optically activeresolving agents (Eliel, E. and Wilen, S. “Stereochemistry of OrganicCompounds,” John Wiley & Sons, Inc., New York, 1994; Lochmuller, C. H.,(1975) J. Chromatogr., 113(3):283-302). Racemic mixtures of chiralcompounds of the invention can be separated and isolated by any suitablemethod, including: (1) formation of ionic, diastereomeric salts withchiral compounds and separation by fractional crystallization or othermethods, (2) formation of diastereomeric compounds with chiralderivatizing reagents, separation of the diastereomers, and conversionto the pure stereoisomers, and (3) separation of the substantially pureor enriched stereoisomers directly under chiral conditions. See: “DrugStereochemistry, Analytical Methods and Pharmacology,” Irving W. Wainer,Ed., Marcel Dekker, Inc., New York (1993).

Under method (1), diastereomeric salts can be formed by reaction ofenantiomerically pure chiral bases such as brucine, quinine, ephedrine,strychnine, α-methyl-β-phenylethylamine (amphetamine), and the like withasymmetric compounds bearing acidic functionality, such as carboxylicacid and sulfonic acid. The diastereomeric salts may be induced toseparate by fractional crystallization or ionic chromatography. Forseparation of the optical isomers of amino compounds, addition of chiralcarboxylic or sulfonic acids, such as camphorsulfonic acid, tartaricacid, mandelic acid, or lactic acid can result in formation of thediastereomeric salts.

Alternatively, by method (2), the substrate to be resolved is reactedwith one enantiomer of a chiral compound to form a diastereomeric pair(E. and Wilen, S. “Stereochemistry of Organic Compounds”, John Wiley &Sons, Inc., 1994, p. 322). Diastereomeric compounds can be formed byreacting asymmetric compounds with enantiomerically pure chiralderivatizing reagents, such as menthyl derivatives, followed byseparation of the diastereomers and hydrolysis to yield the pure orenriched enantiomer. A method of determining optical purity involvesmaking chiral esters, such as a menthyl ester, e.g., (−) menthylchloroformate in the presence of base, or Mosher ester,α-methoxy-α-(trifluoromethyl)phenyl acetate (Jacob III. J. Org. Chem.(1982) 47:4165), of the racemic mixture, and analyzing the ¹H NMRspectrum for the presence of the two atropisomeric enantiomers ordiastereomers. Stable diastereomers of atropisomeric compounds can beseparated and isolated by normal- and reverse-phase chromatographyfollowing methods for separation of atropisomeric naphthyl-isoquinolines(WO 96/15111). By method (3), a racemic mixture of two enantiomers canbe separated by chromatography using a chiral stationary phase (“ChiralLiquid Chromatography” (1989) W. J. Lough, Ed., Chapman and Hall, NewYork; Okamoto, J. Chromatogr., (1990) 513:375-378). Enriched or purifiedenantiomers can be distinguished by methods used to distinguish otherchiral molecules with asymmetric carbon atoms, such as optical rotationand circular dichroism.

General Preparative Procedures

To a microwave reaction tube is charged a4-(2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine compound (1 mmol), aC₆-C₂₀ aryl, C₂-C₂₀ heterocyclyl or C₁-C₂₀ heteroaryl boronic ester (1.3mmol), a 2M solution of sodium carbonate in water (3 mmol),bis(triphenylphosphine)palladium(II) chloride (0.05 mmol) andacetonitrile (200 mmol). The reaction vessel is sealed and the reactionis heated in a Biotage microwave at about 140° C. for 30 minutes. Thereaction mixture is loaded onto a Biotage ISOLUTE SCX-2 column. Thecolumn was first washed with MeOH. The product was eluted with a 2Msolution of ammonia in methanol. After concentration, the crude productis further purified by RP-HPLC. R^(1′) and R^(3′) are R¹ and R³respectively as defined herein, or precursors or protected formsthereof.

The Suzuki-type coupling reaction is useful to attach a heterocycle or aheteroaryl at the 2-position of the pyrimidine ring of a2-chloropyrido[3,2-d]pyrimidine compound. For example,4-(2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine may be combined withabout 1.5 equivalents of4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-c]pyridine,and dissolved in 3 equivalents of sodium carbonate as a 1 molar solutionin water and an equal volume of acetonitrile. A catalytic amount, ormore, of a low valent palladium reagent, such asbis(triphenylphosphine)palladium(II) dichloride, is added. A variety ofboronic acids or boronic esters can be used in place of the indazoleboronic ester indicated. Also alternatively, a nitrogen of the fusedbicyclic heterocycle or a fused bicyclic heteroaryl may be protected,for example as N-THP. In some cases potassium acetate was used in placeof sodium carbonate to adjust the pH of the aqueous layer. The reactionis then heated to about 140-150° C. under pressure in a microwavereactor such as the Biotage Optimizer (Biotage, Inc.) for 10 to 30minutes. The contents are extracted with ethyl acetate, or anotherorganic solvent. After evaporation of the organic layer the4-(2-substitutedpyrido[3,2-d]pyrimidin-4-yl)morpholine compound Suzukicoupling product may be purified on silica or by reverse phase HPLC.

A variety of palladium catalysts can be used during the Suzuki couplingstep to form 4-(2-substitutedpyrido[3,2-d]pyrimidin-4-yl)morpholineexemplary compounds. Low valent, Pd(II) and Pd(0) catalysts may be usedin the Suzuki coupling reaction, including PdCl2(PPh₃)₂, Pd(t-Bu)₃,PdCl₂ dppf CH₂Cl₂, Pd(PPh₃)₄, Pd(OAc)/PPh₃, Cl₂Pd[(Pet₃)]₂, Pd(DIPHOS)₂,Cl₂Pd(Bipy), [PdCl(Ph₂PCH₂PPh₂)]₂, Cl₂Pd[P(o-tol)₃]₂,Pd₂(dba)₃/P(o-tol)₃, Pd₂(dba)/P(furyl)₃, Cl₂Pd[P(furyl)₃]₂,Cl₂Pd(PmePh₂)₂, Cl₂Pd[P(4-F-Ph)₃]₂, Cl₂Pd[P(C₆F₆)₃]₂,Cl₂Pd[P(2-COOH-Ph)(Ph)₂]₂, Cl₂Pd[P(4-COOH-Ph)(Ph)₂]₂, and encapsulatedcatalysts Pd EnCat™ 30, Pd EnCat™ TPP30, and Pd(II)EnCat™ BINAP30 (US2004/0254066).

To a solution of a primary or secondary amine HN(R^(2′))₂ (1 equiv.) andDIEA (about 2 equiv.) in methanol and THF is added4-(6-(bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7(about 1 equiv.). The resulting solution is stirred at room temperaturefor about 1.5 hours. The reaction mixture is evaporated to dryness. Theresidue was dissolved in MeOH and loaded onto an ISOLUTE SCX-2 column.The column is first washed with MeOH. The product is then eluted with a2M solution of ammonia in methanol. After concentration, the crudeproduct is further purified by flash chromatography (0-10% MeOH in DCM).R^(2′) is R² as defined herein, or precursors or protected formsthereof.

To a microwave reaction tube was charged the4-(2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine of interest (1 mmol),the amine of interest (1.5 mmol), cesium carbonate (2 mmol),tris(dibenzylideneacetone)dipalladium (0.05 mmol), XPhos (0.1 mmol) andDMF (220 mmol). The reaction vessel was then sealed and heated in aBiotage microwave at 140° C. for 30 minutes. The reaction mixture wasloaded onto a Biotage ISOLUTE SCX-2 column. The column was first washedwith MeOH. The product was eluted with a 2M solution of ammonia inmethanol. After concentration, the crude product was further purified byRP-HPLC. R^(3′) and R^(5′) are R³ and R⁵ as defined herein, orprecursors or protected forms thereof

To a sealed tube was charged the4-(2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine of interest (1 mmol),the amine of interest (1.05 mmol), potassium phosphate (2.5 mmol),tris(dibenzylideneacetone)dipalladium (0.05 mmol) and XPhos (0.1 mmol)in 1,4-Dioxane (220 mmol). The reaction heated in an oil bath at 150° C.for 2-18 hours. The reaction mixture was then loaded onto a BiotageISOLUTE SCX-2 column. The column was first washed with MeOH. The productwas eluted with a 2M solution of ammonia in methanol. Afterconcentration, the crude product was further purified by RP-HPLC. R^(3′)and R^(5′) are R³ and R⁵ as defined herein, or precursors or protectedforms thereof.

Scheme 1 shows a general synthetic route to pyrido[3,2-d]pyrimidinecompounds via intermediate 6.

EXAMPLES

The chemical reactions described in the Examples may be readily adaptedto prepare a number of other PI3K inhibitors of the invention, andalternative methods for preparing the compounds of this invention aredeemed to be within the scope of this invention. For example, thesynthesis of non-exemplified compounds according to the invention may besuccessfully performed by modifications apparent to those skilled in theart, e.g., by appropriately protecting reactive functional groups, byutilizing other suitable reagents known in the art other than thosedescribed, and/or by making routine modifications of reactionconditions. Alternatively, other reactions disclosed herein or known inthe art will be recognized as having applicability for preparing othercompounds of the invention.

Microwave experiments were carried out using a CEM Explorer, SmithSynthesizer or a Biotage Initiator™, which uses a single-mode resonatorand dynamic field tuning, both of which give reproducibility andcontrol. Temperatures from 40-250° C. can be achieved and pressures upto 20 bar can be reached.

Unless otherwise stated, all reactions were performed under an inert,i.e. argon or nitrogen, atmosphere.

Example 1 5-aminopyrimidine-2,4(1H,3H)-dione 1

Into a 5-L 4-necked round-bottom flask were placed water (2.871 L),ammonia (116.1 mL) and 5-nitropyrimidine-2,4(1H,3H)-dione (180 g, 1.15mol, 1.00 equiv). This was followed by the addition of Na₂S₂O₂ (860 g,6.06 mol, 4.30 equiv) in several batches. The pH value of the solutionwas adjusted to 8 with ammonia (25%). The resulting solution was stirredfor 3 h (hours) at 75° C. The reaction mixture was cooled to 15° C. withan ice/water bath. The solid was collected by filtration. This resultedin 118 g (81%) of 5-aminopyrimidine-2,4(1H,3H)-dione 1 as a yellow solid(see Scheme 1).

Example 2 6-methylpyrido[3,2-d]pyrimidine-2,4(1H,3H)-dione 2

Into a 2000-mL 4-necked round-bottom flask was placed a solution of5-aminopyrimidine-2,4(1H,3H)-dione 1 (217 g, 1.71 mol, 1.00 equiv) inHCl (20%, 1302 mL), then added (E)-but-2-enal (143.5 g, 2.05 mol, 1.20equiv). The resulting solution was heated to reflux for 3 h (hours) inan oil bath. The reaction mixture was cooled and filtered. The filtratewas concentrated under vacuum. The residual solution was diluted with250 mL of water and adjusted to pH 10 with ammonia (25%). The isolatedsolid was collected by filtration, then washed with 2×100 mL of water,2×250 ml of ethanol and 3×500 mL of ether and finally dried in an oven.This resulted in 63 g (21%) of6-methylpyrido[3,2-d]pyrimidine-2,4(1H,3H)-dione 2. ¹H-NMR (400 MHz,CD₃OD, ppm): 7.56 (2H, s), 2.60 (3H, s)

Example 3(2,4-dioxo-1,2,3,4-tetrahydropyrido[3,2-d]pyrimidin-6-yl)methyl acetate3

Into a 5000-mL 4-necked round-bottom flask was placed a solution of6-methylpyrido[3,2-d]pyrimidine-2,4(1H,3H)-dione 2 (120 g, 677.97 mmol,1.00 equiv) in acetic acid (2400 mL). This was followed by the additionof m-CBPA (608 g, 3.51 mol, 5.18 equiv) in several batches. Theresulting solution was stirred overnight at 100° C. The resultingmixture was cooled and concentrated under vacuum. The residue was washedwith 2×1500 mL of ether and 2×500 mL of DCM, then it was dissolved inHOAc (1200 mL) and acetic anhydride (500 mL). The resulting solution wasstirred for 0.5 h at 110° C. The reaction mixture was cooled andfiltered. The filtrate was concentrated under vacuum. The residue waswashed with 2×500 ml of ether and dried. This resulted in 80 g (50%) of(2,4-dioxo-1,2,3,4-tetrahydropyrido[3,2-d]pyrimidin-6-yl)methyl acetate3 as a brown solid.

Example 4 (2,4-dichloropyrido[3,2-d]pyrimidin-6-yl)methyl acetate 4

Into a 1000-mL 3-round-bottom flask was placed a solution of(2,4-dioxo-1,2,3,4-tetrahydropyrido[3,2-d]pyrimidin-6-yl)methyl acetate3 (40 g, 161.70 mmol, 1.00 equiv, 95%) in POCl₃ (400 ml), then addedDIEA (37.3 g, 289.15 mmol, 1.70 equiv). The resulting solution wasstirred for 3 h at 108° C. in an oil bath. The resulting mixture wascooled and concentrated under vacuum. The residue was then quenched bythe addition of 1000 g of ice water. The resulting solution wasextracted with 4×500 mL of dichloromethane. The organic layers werecombined, washed with 2×500 mL of water, dried over sodium sulfate andconcentrated under vacuum. The residue was applied onto a silica gelcolumn and eluted with ethyl acetate/petroleum ether (1:20-1:3). Thisresulted in 24 g (53%) of(2,4-dichloropyrido[3,2-d]pyrimidin-6-yl)methyl acetate 4 as a yellowsolid. ¹H-NMR (400 MHz, CDCl₃, ppm): 8.30-8.32 (1H, d, J=8.8 Hz),7.92-7.94 (1H, d, J=8.8 Hz), 5.47 (2H, s), 2.24 (3H, s)

Example 5 (2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylacetate 5

A mixture of (2,4-dichloropyrido[3,2-d]pyrimidin-6-yl)methyl acetate(1.92 g, 7.06 mmol) according to Srinivasan and Broom (1981) J. Org.Chem. 46:1777-1781, and morpholine (1.3 mL, 14.9 mmol) in ethanol (100mL) was stirred at room temperature for 2 hours. The reaction mixturewas then evaporated to dryness. The crude product was purified by flashchromatography (DCM) to give(2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl acetate 5 (2.2g, 96%). ¹H-NMR (CDCl₃): δ 8.05 (d, 1H), 7.66 (d, 1H), 5.29 (s, 2H), 4.5(s, br, 4H), 3.87 (m, 4H), 2.19 (s, 3H)

Example 6 (2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methanol 6

To a solution of(2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl acetate 5 (2.3g, 7.1 mmol) in THF (50 mL) was added a 1M solution of lithium hydroxidein water (26 mL). The resulting solution was stirred at room temperaturefor 2 hours. The reaction mixture was diluted with water and extractedthree times with dichloromethane. The combined extracts were washed withbrine, dried over sodium sulfate, filtered and concentrated to give(2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methanol 6. LCMS(MH)⁺=281.1. ¹H-NMR (400 MHz, CDCl₃, ppm): 8.09 (1H, d, J=8.8 Hz), 7.66(1H, d, J=8.8 Hz), 4.91 (2H, s), 4.56 (4H, br), 3.91 (4H, t), 2.86 (1H,s)

Example 74-(6-(bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7

(2-Chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methanol 6 (2.0 g,0.0071 mol) and 40 mL dichloromethane were cooled to 0° C. andtriphenylphosphine (2.2 g, 0.0085 mol) was added. The solution wasrecooled to 0° C. and N-Bromosuccinimide NBS (1.5 g, 0.0085 mol) wasadded in one portion. The heterogeneous mixture became homogeneousyellow upon NBS addition. After 30 mins, the progress was checked bytlc. The reaction mixture again became heterogeneous yellow mixture andwas stirred overnight at room temperature. A check by LC/MS showed thereaction was complete. The reaction mixture was concentrated to dryness,taken up in MeOH, and filtered to yield 2.13 gm (87%) of4-(6-(bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7 as ayellow solid. More product 7 precipitated from the mother liquor,filtered, and dried to give a second batch (133 mg) of product 7.Analyzed and confirmed by LC/MS.

Example 82-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol

Following General Procedure B,4-(6-(bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7 and2-(piperidin-4-yl)propan-2-ol were reacted to give2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol.LCMS (MH+)=406.1. ¹H-NMR (DMSO-d₆): δ 8.02 (d, 1H), 7.83 (d, 1H), 4.46(s, br, 4H), 4.01 (s, 1H), 3.78 (m, 4H), 3.68 (s, 2H), 2.88 (m, 2H),1.96 (m, 2H), 1.65 (m, 2H), 1.25 (m, 3H), 1.03 (s, 6H)

Example 1012-(1-((2-(2-methyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol101

Following General Procedure C, 2-methylbenzimidazole and2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 were reacted to give 101. LCMS (MH⁺)=502.3. ¹H-NMR(DMSO-d₆): δ 8.20 (m, 2H), 7.89 (m, 1H), 7.62 (m, 1H), 7.27 (m, 2H),4.56 (s, 4H), 4.08 (s, 1H), 3.84 (m, 4H), 3.73 (s, 2H), 2.92 (m, 2H),2.88 (s, 3H), 1.98 (m, 2H), 1.67 (m, 2H), 1.29 (m, 2H), 1.18 (m, 1H),1.04 (s, 6H)

Example 1022-(1-((2-(2-cyclopropyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol102

Following General Procedure C, 2-cyclopropyl-benzimidazole and2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 were reacted to give 102. LCMS (MH⁺)=528.3. ¹H-NMR(DMSO-d₆): δ 8.19 (d, 1H), 8.03 (m, 1H), 7.89 (d, 1H), 7.44 (m, 1H),7.23 (m, 2H), 4.56 (s, 4H), 4.01 (s, 1H), 3.83 (m, 4H), 3.74 (2, 2H),2.94 (m, 3H), 1.99 (m, 2H), 1.67 (m, 2H), 1.29 (m, 2H), 1.16 (m, 3H),1.08 (m, 2H), 1.04 (s, 6H)

Example 1032-(1-((2-(2-methylbenzofuran-3-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol103

Following General Procedure A,4,4,5,5-tetramethyl-2-(2-methylbenzofuran-3-yl)-1,3,2-dioxaborolane and2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 were reacted to give 103. LCMS (MH⁺)=502.3. ¹H-NMR(DMSO-d₆): δ 8.55 (m, 1H), 8.19 (d, 1H), 7.86 (d, 1H), 7.56 (m, 1H),7.32 (m, 2H), 4.48 (s, 4H), 4.01 (s, 1H), 3.85 (m, 4H), 3.72 (s, 2H),2.96 (s, 3H), 2.93 (m, 2H), 2.01 (m, 2H), 1.66 (m, 2H), 1.29 (m, 2H),1.25 (m, 1H), 1.04 (s, 6H)

Example 1042-ethyl-1-(6-((4-(2-hydroxypropan-2-yl)piperidin-1-yl)methyl)-4-morpholinopyrido[3,2-d]pyrimidin-2-yl)-1H-indazol-3(2H)-one104

Following General Procedure C, 2-ethyl-1H-indazol-3(2H)-one and2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 were reacted to give 104. LCMS (MH⁺)=532.3. ¹H-NMR(DMSO-d₆): δ 8.39 (d, 1H), 8.09 (d, 1H), 7.82 (d, 1H), 7.78 (m, 1H),7.70 (m, 1H), 7.33 (m, 1H), 4.54 (s, 4H), 4.32 (m, 2H), 4.00 (s, 1H),3.85 (m, 4H), 3.68 (s, 2H), 2.90 (m, 2H), 1.97 (m, 2H), 1.64 (m, 2H),1.29 (m, 2H), 1.16 (m, 1H), 1.07 (t, 3H), 1.03 (s, 6H)

Example 1052-(1-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol105

Following General Procedure A,1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indoleand2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 were reacted to give 105. LCMS (MH⁺)=505.2. ¹H-NMR(DMSO-d₆): δ 11.2 (s, 1H), 8.15 (d, 1H), 7.87 (d, 1H), 7.47 (m, 1H),7.44 (s, 1H), 7.01 (m, 1H), 6.75 (s, 1H), 4.49 (s, 4H), 4.02 (s, 1H),3.80 (m, 4H), 3.73 (s, 2H), 2.93 (m, 2H), 1.99 (m, 2H), 1.66 (m, 2H),1.28 (m, 3H), 1.04 (s, 6H)

Example 1062-(1-((2-(5-methyl-1H-pyrazol-3-ylamino)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol106

Following General Procedure C, 5-methyl-1H-pyrazol-3-amine and2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 were reacted to give 106. LCMS (MH⁺)=467.3. ¹H-NMR(DMSO-d₆): δ 11.8 (s, 1H), 8.91 (s, 1H), 7.78 (s, 1H), 7.62 (d, 1H),6.48 (s, 1H), 4.39 (s, 4H), 3.99 (s, 1H), 3.76 (m, 4H), 3.59 (s, 2H),2.90, (m, 2H), 2.19 (s, 3H), 1.93 (m, 2H), 1.63 (m, 2H), 1.24 (m, 2H),1.16 (m, 1H), 1.03 (s, 6H)

Example 1072-(1-((2-(2-aminopyrimidin-5-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol107

Following General Procedure A,5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine and2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 were reacted to give 107. LCMS (MH⁺)=465.3. ¹H-NMR(DMSO-d₆): δ 8.29 (s, 1H), 8.09 (d, 1H), 7.80 (d, 1H), 7.09 (s, 2H),4.50 (s, 4H), 4.01 (s, 1H), 3.81 (m, 4H), 3.68 (s, 2H), 2.91 (m, 2H),1.97 (m, 2H), 1.65 (m, 2H), 1.29 (m, 2H), 1.17 (m, 1H), 1.03 (s, 6H)

Example 108(2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methanol108

(2-Chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methanol 6 (0.1 g) wasreacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 73.4 mg of 108 following reversephase HPLC purification. MS (Q1) 380.1 (M)+

Example 1092-(1-((2-(1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol109

2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 (88 mg) was reacted with 1H-indol-4-ylboronic acid viaGeneral Procedure A to produce 22.8 mg of 109 following reverse phaseHPLC purification. MS (Q1) 487.3 (M)+

Example 1102-(1-((2-(1H-indazol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol110

2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 (88 mg) was reacted with4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole via GeneralProcedure A to produce 29.6 mg of 110 following reverse phase hplcpurification. MS (Q1) 488.3 (M)+

Example 1114-(2-(5-fluoro-1H-indol-4-yl)-6-((3-(tetrahydro-2H-pyran-4-yl)azetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine111

Step 1:4-(2-chloro-6-((3-(tetrahydro-2H-pyran-4-yl)azetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine

4-(6-(Bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7(0.34 g) was reacted with 3-(tetrahydro-2H-pyran-4-yl)azetidine viaGeneral Procedure B to produce crude4-(2-chloro-6-((3-(tetrahydro-2H-pyran-4-yl)azetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine

Step 2: Crude4-(2-chloro-6-((3-(tetrahydro-2H-pyran-4-yl)azetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine(0.1 g) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 49 mg of 111 following reverse phaseHPLC purification. MS (Q1) 503.3 (M)+

Example 1122-(1-((2-(2-isopropyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol112

2-(1-((2-Chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 (0.73 g) was reacted with2-isopropyl-1H-benzo[d]imidazole and sodium tert-butoxide (instead ofcesium carbonate) via General Procedure C to produce 0.192 g of 112following reverse phase HPLC purification. MS (Q1) 530.3 (M)+

Example 1134-(2-(2-isopropyl-1H-benzo[d]imidazol-1-yl)-6-((3-(tetrahydro-2H-pyran-4-yl)azetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine113

4-(2-chloro-6-((3-(tetrahydro-2H-pyran-4-yl)azetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholinefrom Example 111 (0.1 g) was reacted with2-isopropyl-1H-benzo[d]imidazole via General Procedure C to produce 10.2mg of 113 following reverse phase HPLC purification. MS (Q1) 528.3 (M)+

Example 1162-(1-((2-(6-amino-2-methylpyridin-3-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol116

Step 1:6-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine

To 5-bromo-6-methylpyridin-2-amine (0.75 g), bispinacol ester boronate(1.2 eq), potassium acetate (3 eq), Palladium Acetate (0.05 eq) andS-Phos (0.1 eq) in a microwave vial were added Acetonitrile (8 mL) andWater (8 mL). The reaction was heated into a Biotage microwave at 140°C. for 30 minutes. The mixture was diluted with ethyl acetate and washedwith twice with brine. The aqueous layer was back-extracted twice withethyl acetate. The combined organic layers were dried over magnesiumsulfate, filtered and concentrated to yield the crude6-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine

Step 2:2-(1-((2-Chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 (70 mg) was reacted with6-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-aminevia General Procedure A to produce 32.1 mg of 116 following reversephase HPLC purification. MS (Q1) 478.3 (M)+

Example 117 methyl2-(3-((2-(2-isopropyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-1-yl)-2-methylpropanoate117

Step 1: dimethyl(2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylphosphonate

To 4-(6-(bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7(2 g) was added Trimethyl Phosphite (30 eq) and the reaction wasrefluxed at 120° C. for 90 minutes. The reaction was cooled andconcentrated to dryness under vacuum. To the dried crude was addedwater, upon which a yellow solid crashed out and was filtered and driedovernight under vacuum to afford 2 g of pure dimethyl(2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylphosphonate.

Step 2: tert-butyl3-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylene)azetidine-1-carboxylate

To a suspension of dimethyl(2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylphosphonate(0.32 g) in anhydrous THF (3 mL) at 0° C. was added 2.0 M of lithiumdiisopropylamide in tetrahydrofuran (1.4 eq). The resulting solution wasallowed to warm to RT (room temperature) before adding a solution oftert-butyl 3-oxoazetidine-1-carboxylate (1.4 eq) in anhydrous THF (3mL). The reaction mixture was stirred at room temperature for 1 h, thenpartitioned between brine and DCM. The organic layer was isolated, dried(MgSO4) and concentrated in vacuo to give 0.34 g of crude tert-butyl3-((2-chloro-4-morpholinopyrido[3,2-d.]pyrimidin-6-yl)methylene)azetidine-1-carboxylate.

Step 3: tert-butyl3-((2-(2-isopropyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d.]pyrimidin-6-yl)methylene)azetidine-1-carboxylate

To a microwave vial was added tert-butyl3-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylene)azetidine-1-carboxylate,2-isopropyl-1H-benzo[d]imidazole (1.05 eq), sodium tert-butoxide (2 eq),Palladium Acetate (0.1 eq) and Bis(tri-t-butylphosphine)palladium (0.1eq). The tube was flushed with nitrogen for 10 minutes before addingToluene (8 mL). The reaction was microwaved at 145° C. for 20 minutes.The crude was purified by flash column chromatography to afford 0.22 gof tert-butyl3-((2-(2-isopropyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylene)azetidine-1-carboxylate.

Step 4: tert-butyl3-((2-(2-isopropyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidine-1-carboxylate

tert-butyl3-((2-(2-isopropyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylene)azetidine-1-carboxylate(0.2 g) was brought up in MeOH (20 mL) and the flask was purged withnitrogen before addition of 10% Palladium on Carbon (20 mol %). Thereaction was heated at 60° C. for 6 hours, cooled and filtered thrucelite to obtain the crude tert-butyl3-((2-(2-isopropyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidine-1-carboxylatefollowing solvent evaporation.

Step 5: tert-Butyl3-((2-(2-isopropyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidine-1-carboxylate(0.2 g) was deprotected with 4N HCl in dioxane (0.9 mL) at roomtemperature over 1 hour. The reaction was concentrated to dryness toafford crude4-(6-(azetidin-3-ylmethyl)-2-(2-isopropyl-1H-benzo[d]imidazol-1-yl)pyrido[3,2-d]pyrimidin-4-yl)morpholinewhich was reacted with methyl 2-bromo-2-methylpropanoate (3 eq) andpotassium carbonate (6 eq) at 75° C. for 3 hours. The reaction mixtureloaded onto a Biotage isolute cartridge, eluted with 2M NH₃ in MeOH thenpurified by reverse phase HPLC to yield 17.9 mg of 117. MS (Q1) 544.3(M)+

Example 118(2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)(4-(2-hydroxypropan-2-yl)piperidin-1-yl)methanone118

Step 1: 2-chloro-4-morpholinopyrido[3,2-d]pyrimidine-6-carbaldehyde

(2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methanol 6 (1 g) wasreacted with pyridinium chlorochromate (1.1 eq) overnight at roomtemperature in dichloromethane (25 mL). The reaction was filtered thrucelite and run thru a silica plug to get 0.69 g of2-chloro-4-morpholinopyrido[3,2-d]pyrimidine-6-carbaldehyde as a yellowsolid after evaporation.

Step 2: 2-chloro-4-morpholinopyrido[3,2-d]pyrimidine-6-carboxylic acid

2-Chloro-4-morpholinopyrido[3,2-d]pyrimidine-6-carbaldehyde (0.56 g) wasreacted with Oxone (1.1 eq) in DMF (10 mL) overnight. The DMF wasremoved under vacuum and the reaction mixture was brought up into water.The pH was adjusted to 2-3 and product crashed out of solution. Thesolid was collected by filtration and dried to afford 0.45 g of2-chloro-4-morpholinopyrido[3,2-d]pyrimidine-6-carboxylic acid as alight yellow solid.

Step 3:(2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)(4-(2-hydroxypropan-2-yl)piperidin-1-yl)methanone

2-Chloro-4-morpholinopyrido[3,2-d]pyrimidine-6-carboxylic acid (0.12 g)was reacted with 2-(piperidin-4-yl)propan-2-ol (1.2 eq), HATUO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (1.2 eq) and DIPEA (3 eq) in DMF (3.2 mL) for 1hour. The reaction was then extracted with EtOAc and bicarbonatesolution and the organic layer was dried with Magnesium sulfate,filtered and concentrated to give(2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)(4-(2-hydroxypropan-2-yl)piperidin-1-yl)methanonein quantitative yield.

Step 4:(2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)(4-(2-hydroxypropan-2-yl)piperidin-1-yl)methanone(88 mg) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to yield 41.5 mg 118 following reverse phaseHPLC purification. MS (Q1) 519.2 (M)+

Example 119(4-(2-hydroxypropan-2-yl)piperidin-1-yl)(2-(2-isopropyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methanone119

(2-Chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)(4-(2-hydroxypropan-2-yl)piperidin-1-yl)methanonefrom Example 118 (80 mg) was reacted with2-isopropyl-1H-benzo[d]imidazole (1.05 eq), sodium tert-butoxide (2 eq),palladium acetate (0.1 eq) and bis(tri-t-butylphosphine)palladium (0.1eq). The tube was flushed with nitrogen for 10 minutes before addingToluene (2 mL). The reaction was microwaved at 145° C. for 20 minutes.The crude was loaded onto a Biotage isolute scx-2 cartridge, eluted with2M ammonia in MeOH then purified by flash column chromatography toafford 6.6 mg of 119. MS (Q1) 544.3 (M)+

Example 1202-(1-((2-(2-(1,1-difluoroethyl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol120

Step 1:2-(1-((2-(2-aminophenylamino)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol

2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 (0.12 g) was reacted with diaminobenzene via GeneralProcedure C to afford crude2-(1-((2-(2-aminophenylamino)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olafter elution thru a Biotage isolute scx-2 cartridge.

Step 2:2,2-difluoro-N-(2-(6-((4-(2-hydroxypropan-2-yl)piperidin-1-yl)methyl)-4-morpholinopyrido[3,2-d]pyrimidin-2-ylamino)phenyl)propanamide

2-(1-((2-(2-aminophenylamino)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olwas reacted with 2,2-difluoropropanoic acid (2 eq),O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HATU, 1.5 eq) and DIPEA (4 eq) in DMF (4 mL) for 1hr (hour). The reaction mixture was extracted with EtOAc and bicarbonatesolution and the organic layer was dried with Magnesium sulfate,filtered, evaporated and purified via flash column chromatography to get50 mg of2,2-difluoro-N-(2-(6-((4-(2-hydroxypropan-2-yl)piperidin-1-yl)methyl)-4-morpholinopyrido[3,2-d]pyrimidin-2-ylamino)phenyl)propanamide.This intermediate was reacted with acetic acid (1.6 mL) over one week toyield 4.1 mg of 120 following reverse phase HPLC purification. MS (Q1)552.3 (M)+

Example 1212-(4-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-methylpropanamide121

Step 1:2-(4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-methylpropanamide

4-(6-(Bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7 (0.1g) was reacted with 2-methyl-2-(piperazin-1-yl)propanamide via GeneralProcedure B to give 123 mg of2-(4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-methylpropanamide.

Step 2:2-(4-((2-Chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-methylpropanamide(123 mg) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 114 mg of 121 following reverse phaseHPLC purification. MS (Q1) 533.3 (M)+

Example 1222-(1-((2-(6-aminopyridin-3-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol122

2-(1-((2-Chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 (0.1 g) was reacted with 2-aminopyridine-5-boronic acid,pinacol ester via General Procedure A to yield 40.3 mg 122 followingreverse phase HPLC purification. MS (Q1) 464.3 (M)+

Example 1232-(4-((2-(1H-indol-3-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-methylpropanamide123

2-(4-((2-Chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-methylpropanamidefrom Example 121 (115 mg) was reacted with1-(phenylsulfonyl)-3-indoleboronic acid via General Procedure A to yield31.8 mg 123 following phenylsulfonyl group deprotection with aqueouspotassium hydroxide at 50° C. for 2 hours then reverse phase HPLCpurification. MS (Q1) 515.3 (M)+

Example 1242-(4-((2-(6-aminopyridin-3-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-methylpropanamide124

2-(4-((2-Chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-methylpropanamidefrom Example 121 (115 mg) was reacted with 2-aminopyridine-5-boronicacid, pinacol ester via General Procedure A to yield 8.5 mg 124following reverse phase HPLC purification. MS (Q1) 492.3 (M)+

Example 1254-(6-((3-(4,4-difluoropiperidin-1-yl)azetidin-1-yl)methyl)-2-(5-fluoro-1H-indol-4-yl)pyrido[3,2-d]pyrimidin-4-yl)morpholine125

Step 1:4-(2-chloro-6-((3-(4,4-difluoropiperidin-1-yl)azetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine

4-(6-(Bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7 (0.1g) was reacted with 1-(azetidin-3-yl)-4,4-difluoropiperidine via GeneralProcedure B to give 80 mg4-(2-chloro-6-((3-(4,4-difluoropiperidin-1-yl)azetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine.

Step 2:4-(2-chloro-6-((3-(4,4-difluoropiperidin-1-yl)azetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholinewas reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 24.7 mg of 125 following reversephase HPLC purification. MS (Q1) 538.3 (M)+

Example 1264-(1-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholine126

Step 1:4-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholine

tert-butyl 3-morpholinoazetidine-1-carboxylate (31 mg) was treated with4N HCl in dioxane (5 eq) in 1 mL of DCM. The reaction mixture wasstirred for 3 hours at room temperature and then concentrated to drynessto yield 4-(azetidin-3-yl)morpholine in quantitative yield.4-(6-(Bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7(0.413 g) was reacted with 4-(azetidin-3-yl)morpholine via GeneralProcedure B to afford 0.4 g4-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholine

Step 2:4-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholine(119 mg) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 46 mg of 126 following reverse phaseHPLC purification. MS (Q1) 504.2 (M)+

Example 1274-(1-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)piperazin-2-one127

Step 1:4-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)piperazin-2-one

4-(6-(bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7(0.201 g) was reacted with 4-(azetidin-3-yl)piperazin-2-one via GeneralProcedure B to afford 0.18 g4-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)piperazin-2-one.

Step 2:4-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)piperazin-2-one(87 mg) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 46.1 mg of 127 following reversephase HPLC purification. MS (Q1) 517.3 (M)+

Example 1282-(1-((2-(1H-indol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol128

Sodium Hydride (60% oil dispersion, 1.5 eq) was added to a solution ofindole (1.05 eq) in DMF at 0° C. and stirred until gas evolution wascomplete. After 30 minutes, the solution was warmed to room temperatureand2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 (0.15 g) was added. The reaction mixture was microwavedfor 15 minutes at 175° C. and run thru a Biotage isolate scx-2cartridge. The crude was purified by reverse phase HPLC to afford 8.7 mgof 128. MS (Q1) 487.3 (M)+

Example 1294-(2-(5-fluoro-1H-indol-4-yl)-6-((3-(1,1-dioxo)thiomorpholinoazetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine129

Step 1:4-(2-chloro-6-((3-(1,1-dioxo)thiomorpholinoazetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine

4-(6-(Bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7(0.256 g) was reacted with 4-(azetidin-3-yl)1,1-dioxothiomorpholine viaGeneral Procedure B to afford 0.12 g4-(2-chloro-6-((3-(1,1-dioxo)thiomorpholinoazetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine.

Step 2:4-(2-chloro-6-((3-(1,1-dioxo)thiomorpholinoazetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine(0.13 g) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 47.4 mg of 129 following reversephase HPLC purification. MS (Q1) 552.2 (M)+

Example 1304-(2-(5-fluoro-1H-indol-4-yl)-6-((4-(oxetan-3-yl)piperidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine130

Step 1:4-(2-chloro-6-((4-(oxetan-3-yl)piperidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine

4-(6-(Bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7(0.199 g) was reacted with 4-(oxetan-3-yl)piperidine via GeneralProcedure B to afford 0.11 g4-(2-chloro-6-((4-(oxetan-3-yl)piperidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine.

Step 2:4-(2-chloro-6-((4-(oxetan-3-yl)piperidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine(0.11 g) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 59.8 mg of 130 following reversephase HPLC purification. MS (Q1) 503.3 (M)+

Example 1314-(1-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholine131

4-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholinefrom Example 126 (105 mg) was reacted with 2-ethylbenzimidazole andsodium-tert-butoxide (instead of cesium carbonate) via General ProcedureC to afford 15.2 mg of 131 following reverse phase HPLC purification. MS(Q1) 515.3 (M)+

Example 1322-(1-((2-(6-amino-5-methylpyridin-3-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol132

Step 1:3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine

5-bromo-3-methylpyridin-2-amine (1 equiv.), bispinacol ester boronate(1.2 eq), potassium acetate (3 eq), Palladium Acetate (0.05 eq) andS-Phos (0.1 eq) in a microwave vial were added Acetonitrile (8 mL) andWater (8 mL). The reaction was heated into a biotage microwave at 140°C. for 30 minutes. The mixture was diluted with ethyl acetate and washedwith twice with brine. The aqueous layer was back-extracted twice withethyl acetate. The combined organic layers were dried over magnesiumsulfate, filtered and concentrated to yield the crude3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine.

Step 2:2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol(0.185 g) was reacted with3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-aminevia General Procedure A to produce 90.9 mg of 132 following reversephase HPLC purification. MS (Q1) 478.3 (M)+

Example 1332-(1-((2-(1H-indol-3-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol133

2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 (0.1 g) was reacted with1-(phenylsulfonyl)-3-indoleboronic acid via General Procedure A to yield56.9 mg 133 following phenylsulfonyl group deprotection with aqueouspotassium hydroxide at 50° C. for 2 hours then reverse phase HPLCpurification. MS (Q1) 487.3 (M)+

Example 1344-(1-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)piperazin-2-one134

4-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)piperazin-2-onefrom Example 127 (90 mg) was reacted with 2-ethylbenzimidazole andsodium-tert-butoxide (instead of cesium carbonate) via General ProcedureC to afford 10.2 mg of 134 following reverse phase HPLC purification. MS(Q1) 528.3 (M)+

Example 1352-(1-((2-(2-methyl-1H-indol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol135

2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 (0.1 g) was reacted with 2-methylindole andsodium-tert-butoxide (instead of cesium carbonate) via General ProcedureC to afford 19 mg of 135 following reverse phase HPLC purification. MS(Q1) 501.3 (M)+

Example 1362-(1-((2-(2-ethyl-2H-indazol-3-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol136

Step 1:2-(1-((4-morpholino-2-(tributylstannyl)pyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol

A solution of2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 (1.2 g) and bis(tributyltin) (2 eq) in 1,4-dioxane wasdegassed for 15 minutes prior to addition ofbis[di-tert-butyl(4-dimethylaminophenyl)phosphine]dichloropalladium (II)(0.16 eq). The reaction was microwaved at 150° C. for 30 minutes thenpurified by flash column chromatography to afford 1.24 g of2-(1-((4-morpholino-2-(tributylstannyl)pyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olas a clear oil.

Step 2: To a degassed solution of2-(1-((4-morpholino-2-(tributylstannyl)pyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol(0.13 g) and 2-ethyl-3-iodo-2H-indazole (1.05 eq) in 1,4-dioxane (1.5mL) was added copper(I) thiophene-2-carboxylate (1 eq) andtetrakis(triphenylphosphine)palladium(0) (0.05 eq). The reaction wasmicrowaved at 140° C. for 30 minutes and then loaded onto Biotageisolute scx-2 cartridge and eluted with 2M ammonia in Methanol. Thecrude material was purified by reverse phase HPLC to afford 36.6 mg of136. MS (Q1) 516.3 (M)+

Example 137 tert-butyl4-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidine-1-carboxylate137

Step 1: tert-butyl4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylene)piperidine-1-carboxylate

To a suspension of dimethyl(2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylphosphonate fromExample 117 (0.71 g) in anhydrous THF (15 mL) at 0° C. was added 2.0 Mof lithium diisopropylamide in THF (1.4 eq). The resulting solution wasallowed to warm to RT (room temperature) before adding a solution of1-boc-4-piperidone (1.4 eq) in anhydrous THF (3 mL). The reactionmixture was stirred at room temperature for 1 h, then partitionedbetween brine and DCM. The organic layer was isolated, dried (MgSO4),concentrated and recrystallized from methanol. The light yellow solidwas filtered and collected to give 0.72 g of tert-butyl4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylene)piperidine-1-carboxylateas a light yellow solid.

Step 1: tert-butyl4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidine-1-carboxylate

tert-Butyl4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylene)piperidine-1-carboxylate(0.72 g) was brought up in ethanol (80 mL) and ethyl acetate (80 mL) andthe flask was purged with nitrogen before addition of 10% Palladium oncarbon (10 mol %). The reaction was placed under a hydrogen balloon andstirred at room temperature overnight. The reaction was filtered thrucelite and purified by flash column chromatography to obtain 0.55 gtert-butyl4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidine-1-carboxylate.

Step 3: tert-butyl4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidine-1-carboxylate(0.2 g) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A and purified via reverse phase hplc to produce57.5 mg of 137. MS (Q1) 547.3 (M)+

Example 1382-(1-((2-(1H-indazol-3-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol138

To a degassed solution of2-(1-((4-morpholino-2-(tributylstannyl)pyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 136 (0.15 g) and 3-iodo-1H-indazole (1.4 eq) in 1,4-dioxane(1.4 mL) was added Copper(I) thiophene-2-carboxylate (1 eq) andTetrakis(triphenylphosphine)palladium(0) (0.125 eq). The reaction wasmicrowaved at 140° C. for 30 minutes and then loaded onto biotageisolute scx-2 cartridge and eluted with 2M ammonia in Methanol. Thecrude material was purified by reverse phase HPLC to afford 23.4 mg of138. MS (Q1) 488.3 (M)+

Example 1394-(2-(5-fluoro-1H-indol-4-yl)-6-(piperidin-4-ylmethyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine139

Crude tert-butyl4-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidine-1-carboxylate137 was reacted with 4N HCl in dioxane and purified by reverse phaseHPLC to afford 16 mg of 139. MS (Q1) 447.2 (M)+

Example 1401-(((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)(methyl)amino)-2-methylpropan-2-ol140

1-(((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)(methyl)amino)-2-methylpropan-2-olfrom Example 143 (0.12 g) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 35.6 mg of 140 following reversephase HPLC purification. MS (Q1) 465.2 (M)+

Example 1414-(2-(5-fluoro-1H-indol-4-yl)-6-(1-isopropyl-1H-1,2,4-triazol-5-yl)pyrido[3,2-d]pyrimidin-4-yl)morpholine141

Step 1: 2-chloro-4-morpholinopyrido[3,2-d]pyrimidine-6-carboxamide

2-Chloro-4-morpholinopyrido[3,2-d]pyrimidine-6-carboxylic acid fromExample 118 (0.12 g) was reacted with ammonium chloride (7 eq), HATU(1.5 eq) and DIPEA (4 eq) in DMF (5.2 mL) for 30 minutes. The reactionmixture was then diluted with water and the orange solid was collectedby filtration to give 90 mg of2-chloro-4-morpholinopyrido[3,2-d]pyrimidine-6-carboxamide.

Step 2:4-(2-chloro-6-(1-isopropyl-1H-1,2,4-triazol-5-yl)pyrido[3,2-d]pyrimidin-4-yl)morpholine

2-Chloro-4-morpholinopyrido[3,2-d]pyrimidine-6-carboxamide (90 mg) wassuspended in toluene (5 mL) and treated with1,1-dimethoxy-N,N-dimethylmethanamine (11 eq). The reaction was heatedat 95° C. overnight then concentrated to dryness to afford crude(E)-2-chloro-N-((dimethylamino)methylene)-4-morpholinopyrido[3,2-d]pyrimidine-6-carboxamide.This intermediate was subsequently brought up in acetic acid (1.4 mL),isopropyl hydrazine hydrochloride was added and the reaction was stirredat room temperature for 1 hour. The reaction was concentrated to drynessthen extracted with DCM and a saturated bicarbonate solution. Theorganic layer was dried (Mg₂SO₄), filtered and concentrated to give 0.11g crude4-(2-chloro-6-(1-isopropyl-1H-1,2,4-triazol-5-yl)pyrido[3,2-d]pyrimidin-4-yl)morpholine.

Step 3:4-(2-chloro-6-(1-isopropyl-1H-1,2,4-triazol-5-yl)pyrido[3,2-d]pyrimidin-4-yl)morpholine(0.11 g) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 57.7 mg of 141 following reversephase HPLC purification. MS (Q1) 459.2 (M)+

Example 142N-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)tetrahydro-2H-pyran-4-amine142

Step 1:N-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)tetrahydro-2H-pyran-4-amine

4-(6-(bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7 (0.3g) was reacted with tetrahydro-2H-pyran-4-amine via General Procedure Bto afford quantitative yield ofN-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)tetrahydro-2H-pyran-4-amine.

Step 2:N-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)tetrahydro-2H-pyran-4-amine(0.17 g) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 79.1 mg of 142 following reversephase HPLC purification. MS (Q1) 463.2 (M)+

Example 1431-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylamino)-2-methylpropan-2-ol143

Step 1:1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylamino)-2-methylpropan-2-ol

4-(6-(Bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7 (0.3g) was reacted with 1-amino-2-methylpropan-2-ol via General Procedure Bto afford quantitative yield of1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylamino)-2-methylpropan-2-ol.

Step 2:1-(((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)(methyl)amino)-2-methylpropan-2-ol

Paraformaldehyde (20 mg, 1.5 eq) was added to a solution of1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylamino)-2-methylpropan-2-ol(0.16 g) in DCM (2.5 mL). The reaction was stirred for 5 minutes beforeaddition of sodium cyanoborohydride (1.2 eq). Methanol (1 mL) was addedto the reaction mixture which was then stirred overnight at roomtemperature. The reaction was diluted with ethyl acetate and washed withwater. The organic layer was dried over magnesium sulfate, filtered andconcentrated to give 0.12 g of1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)(methyl)amino)-2-methylpropan-2-olas a yellow oil.

Step 3:1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylamino)-2-methylpropan-2-ol(0.16 g) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 75.9 mg of 143 following reversephase HPLC purification. MS (Q1) 451.2 (M)+

Example 1444-(6-((3,3-dimethylpyrrolidin-1-yl)methyl)-2-(5-fluoro-1H-indol-4-yl)pyrido[3,2-d]pyrimidin-4-yl)morpholine144

Step 1:4-(2-chloro-6-((3,3-dimethylpyrrolidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine

4-(6-(Bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7(0.225 g) was reacted with 3,3-dimethylpyrrolidine via General ProcedureB to afford quantitative yield of4-(2-chloro-6-((3,3-dimethylpyrrolidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine.

Step 2:4-(2-chloro-6-((3,3-dimethylpyrrolidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine(0.28 g) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 70.2 mg of 144 following reversephase HPLC purification. MS (Q1) 461.2 (M)+

Example 145N-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)pivalamide145

Step 1: (2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methanamine

To 4-(6-(bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7(0.5 g) in DMF (11 mL) was added sodium azide (2 eq). The reaction washeated at 60° C. for 1 hour and then extracted with ethyl acetate andbrine. The organic layer was dried (Mg₂SO₄), filtered and concentratedto afford 0.437 g of crude4-(6-(azidomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine. Thisintermediate was then suspended in THF (4 mL), triphenylphosphine (2 eq)was added and the reaction was stirred at room temperature for 1.5hours. Water (0.1 mL) was added to the reaction mixture which was thenloaded onto a 10 g biotage isolute scx-2 cartridge then eluted with 2Mammonia in methanol then purified by flash column chromatography toyield 0.27 g(2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methanamine.

Step 2:N-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)pivalamide

(2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methanamine (0.1 g)was added to a pre-stirred (15 minutes) solution of trimethylacetic acid(1.2 eq), 1-hydroxybenzotriazole (1.1 eq.),N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.1 eq.),triethylamine (2 eq.) in N,N-dimethylformamide (2.77 mL). The reactionwas stirred overnight then extracted with ethyl acetate and brine. Theorganic layer was dried (Mg₂SO₄), filtered and concentrated to givequantitative yield ofN-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)pivalamide.

Step 3:N-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)pivalamide(0.13 g) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 44 mg of 145 following reverse phaseHPLC purification. MS (Q1) 463.2 (M)+

Example 1464-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)morpholine146

Step 1:4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)morpholine

4-(6-(Bromomethyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine 7 (0.1g) was reacted with morpholine via General Procedure B to affordquantitative yield of44(2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)morpholine.

Step 2:4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)morpholine(0.1 g) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 94.7 mg of 146 following reversephase HPLC purification. MS (Q1) 449.2 (M)+

Example 147N-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)isobutyramide147

Step 1:N-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)isobutyramide

(2-Chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methanamine fromExample 145 (0.1 g) was added to a pre-stirred (15 minutes) solution of2-methylpropanoic acid (1.2 eq), 1-hydroxybenzotriazole (1.1),N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.1),triethylamine (2 eq) in N,N-dimethylformamide (2.77 mL). The reactionwas stirred overnight then extracted with Ethyl Acetate and brine. Theorganic layer was dried (Mg₂SO₄), filtered and concentrated to givequantitative yield ofN-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)isobutyramide.

Step 2:N-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)isobutyramide(0.125 g) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 14.5 mg of 147 following reversephase HPLC purification. MS (Q1) 449.2 (M)+

Example 1481-(4-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)ethanone148

Step 1:4-(2-chloro-6-(piperidin-4-ylmethyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine

tert-butyl4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidine-1-carboxylatefrom Example 137 (0.33 g) in DCM (5 mL) was treated with 4N HCl indioxane (0.9 mL) for 30 minutes at room temperature. The reactionmixture was concentrated to dryness to afford quantitative yield of4-(2-chloro-6-(piperidin-4-ylmethyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine.

Step 2:1-(4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)ethanone

4-(2-chloro-6-(piperidin-4-ylmethyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine(40 mg) was added to a pre-stirred (15 minutes) solution of acetic acid(5 eq), 1-Hydroxybenzotriazole (HOBt, 1.2 eq.),N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.5 eq.),DIPEA (5 eq) in N,N-Dimethylformamide (DMF, 0.9 mL). The reaction wasstirred overnight then extracted with Ethyl Acetate and brine. Theorganic layer was dried (Mg₂SO₄), filtered and concentrated to givequantitative yield of1-(4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)ethanone.

Step 3:1-(4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)ethanone(30 mg) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 13.7 mg of 148 following reversephase HPLC purification. MS (Q1) 489.2 (M)+

Example 1491-(4-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)-2-methylpropan-1-one149

Step 1:1-(4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)-2-methylpropan-1-one

4-(2-Chloro-6-(piperidin-4-ylmethyl)pyrido[3,2-d]pyrimidin-4-yl)morpholinefrom Example 148 (40 mg) was added to a pre-stirred (15 minutes)solution of 2-methylpropanoic acid (2 eq.), 1-Hydroxybenzotriazole(1.2), N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.5eq.), diisopropylethylamine (DIPEA, 5 eq.) in N,N-Dimethylformamide (0.9mL). The reaction was stirred overnight then extracted with EthylAcetate and brine. The organic layer was dried (Mg₂SO₄), filtered andconcentrated to give quantitative yield of1-(4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)-2-methylpropan-1-one.

Step 2:1-(4-((2-Chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)-2-methylpropan-1-one(60 mg) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 19.9 mg of 149 following reversephase HPLC purification. MS (Q1) 517.3 (M)+

Example 150(S)-4-(1-((2-(2-(1-methoxyethyl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholine150

Following General procedure D,4-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholinefrom Example 126 was reacted with(S)-2-(1-methoxyethyl)-1H-benzo[d]imidazole to give 150. ¹H NMR (400MHz, DMSO) δ 8.18 (d, J=8.6 Hz, 1H), 8.01 (dd, J=6.7, 1.8 Hz, 1H), 7.84(d, J=8.7 Hz, 1H), 7.73 (dd, J=6.7, 2.1 Hz, 1H), 7.38-7.24 (m, 2H), 5.50(q, J=6.4 Hz, 1H), 4.53 (br s, 4H), 3.92-3.79 (m, 6H), 3.63-3.52 (m,4H), 3.45 (t, J=6.2 Hz, 2H), 3.09 (s, 3H), 2.97 (overlapping m, 3H),2.25 (br s, 4H), 1.63 (d, J=6.4 Hz, 3H). LCMS: 345.3

Example 1532-(1-((2-(2-(dimethylamino)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol153

2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 (0.25 g) was reacted withN,N-dimethyl-1H-benzo[d]imidazol-2-amine via General Procedure D toproduce 59.5 mg of 153 following reverse phase HPLC purification. MS(Q1) 531.3 (M)+

Example 1544-(2-(5-fluoro-1H-indol-4-yl)-6-((tetrahydro-2H-pyran-4-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine154

Step 1:4-(6-((2H-pyran-4(3H,5H,6H)-ylidene)methyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine

To a suspension of dimethyl(2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylphosphonate fromExample 117 (0.45 g) in anhydrous THF (6 mL) at 0° C. was added 2.0 M oflithium diisopropylamide in tetrahydrofuran (1.5 eq). The resultingsolution was allowed to warm to RT before adding a solution ofdihydro-2H-pyran-4(3H)-one (1.5 eq) in anhydrous THF (3 mL). Thereaction mixture was stirred at room temperature for 1 h, thenpartitioned between brine and Ethyl Acetate. The organic layer wasisolated, dried (MgSO₄), concentrated and recrystallized from methanol.The light yellow solid was filtered and collected to give 0.36 g of4-(6-((2H-pyran-4(3H,5H,6H)-ylidene)methyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholineas a light yellow solid.

Step 2:4-(2-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine

4-(6-((2H-Pyran-4(3H,5H,6H)-ylidene)methyl)-2-chloropyrido[3,2-d]pyrimidin-4-yl)morpholine(0.22 g) was brought up in ethanol (30 mL) and ethyl acetate (30 mL) andthe flask was purged with nitrogen before addition of 10% Palladium onCarbon (10 mol %). The reaction was placed under a hydrogen balloon andstirred at room temperature for 2.5 hours. The reaction was filteredthru celite and purified by flash column chromatography to obtain 0.11 g4-(2-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine.

Step 3:4-(2-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine(0.05 g) was reacted with1-(tert-butyldimethylsilyl)-5-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indolevia General Procedure A to produce 38.8 mg of 154 following reversephase HPLC purification. MS (Q1) 448.2 (M)+

Example 1562-(1-((4-morpholino-2-(2-(trifluoromethyl)-1H-benzo[d]imidazol-1-yl)pyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol156

Following General Procedure D,2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 was reacted with 2-(trifluoromethyl)-1H-benzo[d]imidazoleto give 156. ¹H NMR (400 MHz, DMSO) δ 8.20 (d, J=8.6 Hz, 1H), 8.14 (d,J=8.3 Hz, 1H), 7.94 (d, J=8.6 Hz, 2H), 7.56 (t, J=7.5 Hz, 1H), 7.49 (t,J=7.6 Hz, 1H), 4.56 (s, 4H), 4.04 (s, 1H), 3.86-3.79 (m, 4H), 3.75 (s,2H), 2.93 (d, J=11.0 Hz, 2H), 2.00 (t, J=10.9 Hz, 2H), 1.67 (d, J=12.1Hz, 2H), 1.38-1.12 (m, 3H), 1.04 (s, 6H). LCMS: 556.3

Example 1572-(1-((2-(2-(difluoromethyl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol157

Following General Procedure D,2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 was reacted with 2-(difluoromethyl)-1H-benzo[d]imidazoleto give 157. ¹H NMR (400 MHz, DMSO) δ 8.43 (d, J=8.3 Hz, 1H), 8.23 (d,J=8.7 Hz, 1H), 8.00-7.72 (overlapping m, 3H), 7.51 (app t, J=7.7 Hz,1H), 7.44 (app t, J=7.7 Hz, 1H), 4.56 (br s, 4H), 4.03 (s, 1H), 3.86 (brs, 4H), 3.74 (br s, 2H), 2.92 (d, J=10.6 Hz, 2H), 1.99 (t, J=11.3 Hz,2H), 1.67 (d, J=11.8 Hz, 2H), 1.38-1.10 (overlapping m, 3H), 1.04 (s,6H). LCMS: 538.3

Example 1582-(1-(6-((4-(2-hydroxypropan-2-yl)piperidin-1-yl)methyl)-4-morpholinopyrido[3,2-d]pyrimidin-2-yl)-1H-benzo[d]imidazol-2-yl)acetonitrile158

Following General Procedure D,2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 was reacted with 2-(1H-benzo[d]imidazol-2-yl)acetonitrileto give 158. ¹H NMR (400 MHz, DMSO) δ 8.32 (d, J=7.8 Hz, 1H), 8.21 (d,J=8.5 Hz, 1H), 7.91 (d, J=8.6 Hz, 1H), 7.76 (d, J=7.8 Hz, 1H), 7.38(overlapping m, 2H), 4.88 (s, 2H), 4.54 (s, 4H), 4.04 (s, 1H), 3.86 (s,4H), 3.73 (s, 2H), 2.92 (d, J=10.5 Hz, 2H), 1.99 (t, J=11.5 Hz, 2H),1.67 (d, J=11.7 Hz, 2H), 1.26 (overlapping m, 3H), 1.04 (s, 6H). LCMS:527.3

Example 163N,N-dimethyl-1-(4-morpholino-6-((3-(1,1-dioxo)thiomorpholinoazetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-2-yl)-1H-benzo[d]imidazol-2-amine163

4-(2-chloro-6-((3-(1,1-dioxo)thiomorpholinoazetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholinefrom Example 129 (0.1 g) was reacted withN,N-dimethyl-1H-benzo[d]imidazol-2-amine via General Procedure D toproduce 12.5 mg of 163 following reverse phase HPLC purification. MS(Q1) 578.2 (M)+

Example 164(S)-1-(4-((2-(2-(1-methoxyethyl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)ethanone164

1-(4-((2-Chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)ethanonefrom Example 148 (0.125 g) was reacted with(S)-2-(1-methoxyethyl)-1H-benzo[d]imidazole via General Procedure D toproduce 22.1 mg of 164 following reverse phase HPLC purification. MS(Q1) 530.3 (M)+

Example 1651-(4-((2-(2-(dimethylamino)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)ethanone165

1-(4-((2-Chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)ethanonefrom Example 148 (0.125 g) was reacted withN,N-dimethyl-1H-benzo[d]imidazol-2-amine via General Procedure D toproduce 47.5 mg of 165 following reverse phase HPLC purification. MS(Q1) 515.2 (M)+

Example 1662-(1-((2-([1,2,4]triazolo[1,5-a]pyridin-5-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol166

2-(1-((2-Chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 (50 mg, 0.12 mmol) was dissolved in 2 mL of THF andtreated with CuI (1.2 mg, 0.006 mmol) and5-(tributylstannyl)-[1,2,4]triazolo[1,5-a]pyridine (65.4 mg, 0.16 mmol).The solution was sparged with bubbling nitrogen andtetrakis(triphenylphosphine)palladium (0) (7 mg, 0.006 mmol) was added.The mixture was heated in an Agilent microwave (140° C., 20 min). Thesolution was cooled to room temperature and concentrated. Purificationby reverse phase HPLC gave 166 as a colorless solid. ¹H NMR (400 MHz,DMSO) δ 8.52 (s, 1H), 8.21 (d, J=8.6 Hz, 1H), 7.99 (d, J=8.9 Hz, 1H),7.91 (d, J=8.7 Hz, 1H), 7.84-7.74 (m, 1H), 7.65 (d, J=7.0 Hz, 1H), 4.51(s, 4H), 4.03 (s, 1H), 3.83-3.77 (m, 4H), 3.75 (s, 2H), 2.93 (d, J=11.1Hz, 2H), 2.00 (t, J=11.2 Hz, 2H), 1.67 (d, J=11.4 Hz, 2H), 1.24(overlapping m, 3H), 1.04 (s, 6H). LCMS: 489.2

Example 167N,N-dimethyl-1-(4-morpholino-6-((3-morpholinoazetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-2-yl)-1H-benzo[d]imidazol-2-amine167

4-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholinefrom Example 126 (75 mg) was reacted withN,N-dimethyl-1H-benzo[d]imidazol-2-amine via General Procedure D toproduce 6.8 mg of 167 following reverse phase HPLC purification. MS (Q1)530.2 (M)+

Example 1682-(1-((2-(2-(2-hydroxyethyl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol168

2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 (0.15 g) was reacted with2-(1H-benzo[d]imidazol-2-yl)ethanol via General Procedure D to produce19 mg of 168 following reverse phase HPLC purification. MS (Q1) 532.3(M)+

Example 1694-(2-(2-(difluoromethyl)-1H-benzo[d]imidazol-1-yl)-6-((3-(1,1-dioxo)thiomorpholinoazetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine169

4-(2-chloro-6-((3-(1,1-dioxo)thiomorpholinoazetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholinefrom Example 129 (0.125 g) was reacted with2-(difluoromethyl)-1H-benzo[d]imidazole via General Procedure D toproduce 50.9 mg of 169 following reverse phase HPLC purification. MS(Q1) 585.2 (M)+

Example 1711-(4-((2-(2-(difluoromethyl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)ethanone171

1-(4-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)ethanonefrom Example 148 (0.135 g) was reacted with2-(difluoromethyl)-1H-benzo[d]imidazole via General Procedure D toproduce 31.4 mg of 171 following reverse phase HPLC purification. MS(Q1) 522.2 (M)+

Example 1724-(1-((2-(2-(difluoromethyl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholine172

4-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholinefrom Example 126 (0.07 g) was reacted with2-(difluoromethyl)-1H-benzo[d]imidazole via General Procedure D toproduce 10.7 mg of 172 following reverse phase HPLC purification. MS(Q1) 537.2 (M)+

Example 1732-(1-((4-morpholino-2-(2-(2,2,2-trifluoroethyl)-1H-benzo[d]imidazol-1-yl)pyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol173

A small vial was charged with2-(1-((2-chloro-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-olfrom Example 8 (0.070 g, 0.17 mmol),2-(2,2,2-trifluoro-ethyl)-1H-benzoimidazole (0.042 g, 0.21 mmol), XPhos(0.012 g, 0.026 mmol) and Potassium phosphate (0.110 g, 0.519 mmol). Dry1,4-dioxane (1.9 mL, 0.024 mol) was added. The whole was sparged withnitrogen for 3 min. Tris(dibenzylideneacetone)dipalladium(0) (0.012 g,0.013 mmol) was added under nitrogen. The vial was sealed and heated at155° C. (heating block temp) for 4 h. LC/MS indicates complete reaction.The solution was diluted with water and extracted with dichloromethane 3times. The combined organics were washed with brine and dried oversodium sulfate. The material was purified by reverse phase HPLC to give51.3 mg (53% yield) of 173 as a white solid. LCMS (MH⁺)=570.3. ¹H NMR(400 MHz, DMSO) δ 8.25-8.15 (m, 2H), 7.91 (d, J=8.7 Hz, 1H), 7.76 (d,J=7.5 Hz, 1H), 7.42-7.31 (m, 2H), 4.65 (q, J=10.7 Hz, 2H), 4.90-4.25 (brm, 4H), 4.05 (s, 1H), 3.88-3.80 (m, 4H), 3.74 (s, 2H), 2.92 (d, J=11.1Hz, 2H), 1.99 (t, J=11.1 Hz, 2H), 1.67 (d, J=11.2 Hz, 2H), 1.37-1.23 (m,2H), 1.22-1.12 (m, 1H), 1.04 (s, 6H)

Example 901 PI3K Isoform Inhibition Assay (p110 alpha, beta, gamma,delta: α, β, γ, δ)

PI3K enzymatic activity was assayed by measuring the amount of productphosphatidylinositol 3,4,5-phosphate (PIP3) formed from substrate 4,5phosphatidylinositol 4,5-phosphate (PIP2) using a fluorescencepolarization displacement assay. The decrease in fluorescencepolarization of a fluorescent PIP₃ probe is measured as it is displacedfrom a PIP₃-binding protein GRP-1 detector by PI3K-catalyzed product.Assays were conducted in 384-well black Proxiplates in the presence of10 mM Tris (pH 7.5), 50 mM NaCl, 4 mM MgCl₂, 5% glycerol, 25 μM ATP, 10μM PIP₂ (Echelon Biosciences), 0.05%3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, 1 mMdithiothreitol, and 2% DMSO. The kinase reactions were initiated by theaddition of 40 ng/mL p110α/p85α, 300 ng/mL p110β/p85α, 40 ng/mL p110γ,or 40 ng/mL p110δ/p85α(Upstate Group, Millipore; Dundee, UK), and 10 μMPIP₂ (Echelon Biosciences) to the wells. The reactions were stopped attime points that yielded a fixed change in fluorescence polarizationconsistent with initial rate conditions (typically 30 minutes), by theaddition of 12.5 mM EDTA, 100 nM GRP-1 detector, and 5 nMtetramethylrhodamine-labeled PIP₃ (TAMRA-PIP₃; Echelon Biosciences).After 60 minutes of incubation at room temperature to allowequilibration of labeled and unlabeled PIP3 binding, the parallel andperpendicular components of the fluorescence emissions from each samplewere measured at an excitation wavelength of 530 nm and an emissionwavelength of 590 nm using an Envision fluorescent plate reader with arhodamine filter (PerkinElmer Life and Analytical Sciences; Wellesley,Mass.). The assay is capable of detecting 0.1-2.0 μM PIP₃ product. TheIC₅₀ values were obtained by fitting the dose-dependent inhibition datato a 4-parameter equation using Assay Explorer software (MDL, San Ramon,Calif.).

Alternatively, inhibition of PI3K was determined in a radiometric assayusing purified, recombinant enzyme and ATP at a concentration of 1 μM.The Formula I compound was serially diluted in 100% DMSO. The kinasereaction was incubated for 1 h at room temperature, and the reaction wasterminated by the addition of PBS. IC₅₀ values were subsequentlydetermined using sigmoidal dose-response curve fit (variable slope).

The same protocol may be used to establish IC₅₀ values for p110α (alpha)PI3K binding.

Recombinant PI3K p110 isoforms alpha, beta, and delta may be preparedand purified according to US 2008/0275067 from recombinant PI3Kheterodimeric complexes consisting of a p110 catalytic subunit and a p85regulatory subunit overexpressed using the BAC-TO-BAC® HT baculovirusexpression system (GIBCO/BRL), and then purified for use in biochemicalassays. The four Class I PI 3-kinases are cloned into baculovirusvectors as follows:

p110 delta: A FLAG™-tagged (Eastman Kodak Co., U.S. Pat. No. 4,703,004;U.S. Pat. No. 4,782,137; U.S. Pat. No. 4,851,341) version of humanp110.delta (Chantry et al., J. Biol. Chem. (1997) 272:19236-41) issubcloned using standard recombinant DNA techniques into the BamH1-Xba1site of the insect cell expression vector pFastbac HTb (LifeTechnologies, Gaithersburg, Md.), such that the clone is in frame withthe His tag of the vector.

p110 alpha: Similar to the method used for p110 delta, described above,a FLAG™-tagged version of p110 alpha (Volinia et al (1994) Genomics,24(3):427-77) was subcloned in BamH1-HindIII sites of pFastbac HTb (LifeTechnologies) such that the clone was in frame with the His tag of thevector.

p110 beta: A p110 beta (see Hu et al (1993) Mol. Cell. Biol.,13:7677-88) clone was amplified from the human MARATHON™ Ready spleencDNA library (Clontech, Palo Alto Calif.) according to themanufacturer's protocol using the specified primers.

The p110 delta binding IC50 values and delta/alpha selectivity ofselected compounds from Table 1 include:

p110 delta IC50 IC50 p110 alpha/IC50 Compound No. (micromolar) p110delta 101 0.0125 33 102 0.00794 71 103 0.0105 200 104 0.0118 83 1050.00343 >350 106 0.0669 31 107 0.00257 5.4 108 0.0265 13.5 1090.00282 >350 110 0.00632 21 111 0.00532 281 112 0.00533 44 113 0.0085228.4 114 0.00178 53 115 0.00711 58 116 0.00933 43 117 0.00509 37 1180.013 77 119 0.0878 15 120 0.00231 45 121 0.00129 203 122 0.00542 15 1230.00448 55 124 0.00203 14 125 0.00152 246 126 0.00114 257 127 0.00175141 128 0.0819 26 129 0.000845 440 130 0.00312 352 131 0.00223 36 1320.0224 10 133 0.0238 22 134 0.00356 10.9 135 0.0256 82 141 0.0636 9.4143 0.0359 58 156 0.00286 42 159 0.873 1.4 160 0.0983 21 161 0.118 17.8188 0.00869 0.18 197 0.00572 0.20 198 0.00388 0.27 199 0.728 0.95 2000.716 0.97 201 0.00491 0.34

Example 902 Collagen Induced Arthritis Efficacy Test

The efficacy of Formula I compound inhibitors of PI3K delta to inhibitthe induction and/or progression of collagen induced arthritis wastested in mice. DBA1/J male mice (Jackson Labs; 5-6 weeks of age) areacclimatized for one week and are then injected intra-dermally at thebase of the tail with 0.1 ml of an emulsion of Bovine Type II Collagen(100 mg) and an equal volume of Complete Freunds Adjuvant (200 mgMycobacterium tuberculosis). Three weeks later, mice are injectedintra-dermally at the base of the tail with 0.1 ml of an emulsion ofBovine Type II Collagen (100 mg) and an equal volume of IncompleteFreunds Adjuvant for boost. Dosing generally starts as soon as animalsdisplay signs of joint inflammation or clinical score 1-2.

All mice are evaluated 2-3 times a week for arthritis using amacroscopic scoring system for each paw. At the end of the experimentclinical scores are obtained to evaluate the intensity of edema in thefour paws. A score of 0 to 4 is assigned to each paw. Animals are scored0 when no inflammatory signs (swelling and redness) are observed in anyof the small joints (intraphalangeal, metacarpophalangeal,metatarsophalangeal) or large joints (wrist/carpus, ankle/tarsus).Animals are scored 1 when very slight to slight inflammation wasobserved (swelling and/or redness of paw or one digit), 2 moderateedema(swelling in two or more joint), 3 severe edema(gross swelling ofthe paw with more than two joints involved), and 4 when very severeedema(severe arthritis of the entire paw and digits) is present. Thearthritic index for each mouse is evaluated by adding the four scores ofthe individual paws, giving a maximum score of 16. Plasma and serumsamples are taken at 1 hour (orbital bleed) post dose and 24 hrs(cardiac puncture) post dose. Samples are stored at −20° C. untilanalysis. At termination, the hind paws are transected at the distaltibia, just proximal to the tarsal joint. The left and right hind pawsare placed in the histology cassettes individually and fixed in 10%formalin. These paws are sent to histology dept for further process.

Materials: Bovine Type II collagen, immunization grade, 2 mg/ml (5ml/vial) in 0.05 M acetic acid (solution), store at −20° C., fromChondrex, LLC, Seattle, Wash. Adjuvant complete H37 Ra, 6×10 ml/box,contains 1 mg/ml Mycobacterium tuberculosis. For use in animalimmunological studies, for laboratory use, store at +4° C., from DifcoLaboratories, Detroit, Mich. 48232-7058 USA. Adjuvant Incomplete H37 Ra,6×10 ml/box: For use in animal immunological studies, for laboratoryuse, store at +4° C., from Difco Laboratories.

Example 903 CD69 Whole Blood Assay

Human blood is obtained from healthy volunteers, with the followingrestrictions: 1 week drug-free, non-smokers. Blood (approximately 20 mlsto test 8 compounds) is collected by venipuncture into Vacutainer tubeswith sodium heparin.

Cynomolgus monkey blood is obtained courtesy of the LAT group frommonkeys not previously exposed to, or after a washout period from,chemical dosing. Additional cyno blood draws may be collected during thecourse of pharmacokinetic or toxicology studies. Blood (25-30 mls fornaïve monkeys or 3-4 mls from monkeys on studies requiring repeateddraws) is collected by venipuncture into Vacutainer tubes with sodiumheparin.

Solutions of Formula I compounds at 1000 or 2000 μM in PBS (20×), arediluted by three-fold serial dilutions in 10% DMSO in PBS for a ninepoint dose-response curve. An aliquot of 5.5 μl of each compound isadded in duplicate to a 2 ml 96-well plate; 5.5 μl of 10% DMSO in PBS isadded as control and no-stimulus wells. Human whole blood—HWB (100 μl)is added to each well. After mixing the plates are incubated at 37° C.,5% CO₂, 100% humidity for 30 minutes. Goat F(ab′)2 anti-human IgM (10 μlof a 500 μg/ml solution, 50 μg/ml final) is added to each well (exceptthe no-stimulus wells) with mixing and the plates are incubated for anadditional 20 hours. At the end of the 20 hour incubation, samples areincubated with florescent labeled antibodies for 30 minutes, at 37° C.,5% CO₂, 100% humidity. Include induced control, unstained and singlestains for compensation adjustments and initial voltage settings.Samples are then lysed with Pharmingen Lyse according to themanufacturer's instructions. Samples are then transferred to a 96 wellplate suitable to be run on the AMS 96 well system on the BD CaliburFACs machine. Data acquired and Mean Fluorescence Intensity values wereobtained using Cell Quest Software. Results are initially analyzed byFACS analysis software (Flow Jo). The IC50 for test compounds is definedas the concentration which decreases by 50% the percent positive of CD69cells that are also CD20 positive stimulated by anti-IgM (average of 8control wells, after subtraction of the average of 8 wells for theno-stimulus background). The IC50 values are calculated by Activityl)aseusing Xlfit version 3, equation 201.

The IC50 values of selected compounds from Table 1 in the CD69 WholeBlood Assay include:

Compound No. IC50 (micromolar) 105 0.0412 112 0.0838 114 0.012

The foregoing description is considered as illustrative only of theprinciples of the invention. Further, since numerous modifications andchanges will be readily apparent to those skilled in the art, it is notdesired to limit the invention to the exact construction and processshown as described above. Accordingly, all suitable modifications andequivalents may be considered to fall within the scope of the inventionas defined by the claims that follow.

The words “comprise,” “comprising,” “include,” “including,” and“includes” when used in this specification and in the following claimsare intended to specify the presence of stated features, integers,components, or steps, but they do not preclude the presence or additionof one or more other features, integers, components, steps, or groupsthereof.

We claim:
 1. A compound selected from Formula I:

and stereoisomers, geometric isomers, tautomers, or pharmaceuticallyacceptable salts thereof, wherein R¹ is selected from F, Cl, Br, I,N(R²)₂, OR², SR², SOR², SO₂R², SO₂N(R²)₂, C₁-C₁₂ alkyl, C₂-C₈ alkenyl,C₂-C₈ alkynyl, C₆-C₂₀ aryl, C₃-C₁₂ carbocyclyl, C₂-C₂₀ heterocyclyl,C₁-C₂₀ heteroaryl, —(C₁-C₁₂ alkylene)-(C₃-C₁₂ carbocyclyl), —(C₁-C₁₂alkylene)-(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂ alkylene)-(C₂-C₂₀heterocyclyl)-(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂ alkylene)-(C₂-C₂₀heterocyclyl)-(C₃-C₁₂ carbocyclyl), —(C₁-C₁₂ alkylene)-(C₂-C₂₀heterocyclyl)-C(═O)—(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂ alkylene)-(C₁-C₂₀heteroaryl), —(C₁-C₁₂ alkylene)-(C₂-C₂₀ heterocyclyl)-(C₁-C₁₂ alkyl),—(C₁-C₁₂ alkylene)-(C₆-C₂₀ aryl)-(C₁-C₁₂ alkyl), —(C₁-C₁₂alkylene)-(C₁-C₂₀ heteroaryl)-(C₁-C₁₂ alkyl), —(C₁-C₁₂alkylene)-C(═O)—(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂ alkylene)-N(R²)₂,—(C₁-C₁₂ alkylene)-NR²C(═O)R², —(C₁-C₁₂ alkylene)-NR²—(C₁-C₁₂ alkyl),—(C₁-C₁₂ alkylene)-N(C₁-C₁₂ alkyl)(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂alkylene)-NR²—(C₁-C₁₂ alkylene)-(C₁-C₂₀ heteroaryl), —(C₁-C₁₂alkylene)-NR²—(C₁-C₁₂ alkylene)-(C₁-C₂₀ heterocyclyl), —(C₁-C₁₂alkylene)-NR²—(C₁-C₁₂ alkylene)-NHC(═O)—(C₁-C₂₀ heteroaryl), —(C₁-C₁₂alkylene)-(C₂-C₂₀ heterocyclyl)-N(C₁-C₁₂ alkyl)R², —(C₁-C₁₂alkylene)-(C₂-C₂₀ heterocyclyl)-(C₁-C₁₂ alkyl)-N(C₁-C₁₂ alkyl)R²,—(C₁-C₁₂ alkylene)-NR²—(C₂-C₂₀ heterocyclyl), —(C₂-C₁₂alkenylene)-(C₂-C₂₀ heterocyclyl), —(C₂-C₂₀ heterocyclyl)-(C₁-C₁₂alkyl), —NR²—(C₂-C₂₀ heterocyclyl), —C(═O)—(C₂-C₂₀ heterocyclyl), and—C(═O)—(C₁-C₁₂ alkyl), where alkyl, alkenyl, alkynyl, alkylene,carbocyclyl, heterocyclyl, aryl, and heteroaryl are optionallysubstituted with one or more groups independently selected from F, Cl,Br, I, —CH₃, —CH₂CH₃, —CH₂CH(CH₃)₂, —CH₂OH, —CH₂CH₂OH, —C(CH₃)₂OH,—CH(OH)CH(CH₃)₂, —C(CH₃)₂CH₂OH, —CH₂CH₂SO₂CH₃, —CN, —CF₃, —CHF₂, —CO₂H,—COCH₃, —CO₂CH₃, —C(CH₃)₂CO₂CH₃, —CO₂C(CH₃)₃, —COCH(OH)CH₃, —COCH(CH₃)₂,CONH₂, —CONHCH₃, —CON(CH₃)₂, —C(CH₃)₂CONH₂, —NO₂, —NH₂, —NHCH₃,—N(CH₃)₂, —NHCOCH₃, —N(CH₃)COCH₃, —NHS(O)₂CH₃, —N(CH₃)C(CH₃)₂CONH₂,—N(CH₃)CH₂CH₂S(O)₂CH₃, ═O, —OH, —OCH₃, —S(O)₂N(CH₃)₂, —SCH₃, —CH₂OCH₃,—S(O)₂CH₃, cyclopropyl, oxetanyl, and morpholino; R² is H; R³ isselected from

each of which are optionally substituted with one or more groupsindependently selected from F, Cl, Br, I, —CH₃, —CH₂CH₃, —CH₂CH₂CH₃,—CH(CH₃)₂, —C(CH₃)₃, —CH₂OCH₃, —CHF₂, —CN, —CF₃, —CH₂OH, —CH₂OCH₃,—CH₂CH₂OH, —CH₂C(CH₃)₂OH, —CH(CH₃)OH, —CH(CH₂CH₃)OH, —CH₂CH(OH)CH₃,—CH₂CH(OCH₃)CH₃, —C(CH₃)₂OH, —C(CH₃)₂OCH₃, —CH(CH₃)F, —C(CH₃)F₂,—CH(CH₂CH₃)F, —C(CH₂CH₃)₂F, —CO₂H, —CONH₂, —CON(CH₂CH₃)₂, —COCH₃,—CON(CH₃)₂, —NO₂, —NH₂, —NHCH₃, —N(CH₃)₂, —NHCH₂CH₃, —NHCH(CH₃)₂,—NHCH₂CH₂OH, —NHCH₂CH₂OCH₃, —NHCOCH₃, —NHCOCH₂CH₃, —NHCOCH₂OH,—NHS(O)₂CH₃, —N(CH₃)S(O)₂CH₃, ═O, —OH, —OCH₃, —OCH₂CH₃, —OCH(CH₃)₂, —SH,—NHC(═O)NHCH₃, —NHC(═O)NHCH₂CH₃, —S(O)CH₃, —S(O)CH₂CH₃, —S(O)₂CH₃,—S(O)₂NH₂, —S(O)₂NHCH₃, —S(O)₂N(CH₃)₂, —CH₂S(O)₂CH₃, and a groupselected from

R⁴ is H; R⁵ is selected from H, C₁-C₁₂ alkyl, C₂-C₈ alkenyl, C₂-C₈alkynyl, —(C₁-C₁₂ alkylene)-(C₃-C₁₂ carbocyclyl), —(C₁-C₁₂alkylene)-(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂ alkylene)-C(═O)—(C₂-C₂₀heterocyclyl), —(C₁-C₁₂ alkylene)-(C₆-C₂₀ aryl), and —(C₁-C₁₂alkylene)-(C₁-C₂₀ heteroaryl), where alkyl, alkenyl, alkynyl, alkylene,carbocyclyl, heterocyclyl, aryl, and heteroaryl are optionallysubstituted with one or more groups independently selected from F, Cl,Br, I, —CH₃, —CH₂OH, —CN, —CF₃, —CO₂H, —COCH₃, —CO₂CH₃, —CONH₂,—CONHCH₃, —CON(CH₃)₂, —NO₂, —NH₂, —NHCH₃, —NHCOCH₃, —NHS(O)₂CH₃, —OH,—OCH₃, —S(O)₂N(CH₃)₂, —SCH₃, —CH₂OCH₃, and —S(O)₂CH₃; and n is 0 or 1.2. The compound of claim 1 wherein R¹ is selected from the structures

where the wavy line indicates the site of attachment.
 3. The compound ofclaim 1 wherein R³ is selected from:

where the wavy line indicates the site of attachment and R¹⁴ is selectedfrom F, Cl, Br, I, —CH₃, —CN, —CF₃, —CH₂OH, —CO₂H, —CONH₂, —CON(CH₃)₂,—NO₂, —NH₂, —NHCH₃, —NHCOCH₃, —OH, —OCH₃, —SH, —NHC(═O)NHCH₃,—NHC(═O)NHCH₂CH₃, and —S(O)₂CH₃.
 4. The compound of claim 1 wherein R³is 1H-indazol-4-yl and n is
 0. 5. The compound of claim 1 selected from2-(1-((2-(2-methyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;2-(1-((2-(2-cyclopropyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;2-(1-((2-(2-methylbenzofuran-3-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;2-ethyl-1-(6-((4-(2-hydroxypropan-2-yl)piperidin-1-yl)methyl)-4-morpholinopyrido[3,2-d]pyrimidin-2-yl)-1H-indazol-3(2H)-one;2-(1-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;(2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methanol;2-(1-((2-(1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;2-(1-((2-(1H-indazol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;4-(2-(5-fluoro-1H-indol-4-yl)-6-(3-(tetrahydro-2H-pyran-4-yl)azetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine;2-(1-((2-(2-isopropyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;4-(2-(2-isopropyl-1H-benzo[d]imidazol-1-yl)-6-43-(tetrahydro-2H-pyran-4-yl)azetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine;(S)-2-(1-((2-(2-(1-methoxyethyl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;(R)-2-(1-((2-(2-(1-methoxyethyl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;methyl2-(3-((2-(2-isopropyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-1-yl)-2-methylpropanoate;(2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)(4-(2-hydroxypropan-2-yl)piperidin-1-yl)methanone;(4-(2-hydroxypropan-2-yl)piperidin-1-yl)(2-(2-isopropyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methanone;2-(1-((2-(2-(1,1-difluoroethyl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;2-(4-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-methylpropanamide;2-(4-((2-(1H-indol-3-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-methylpropanamide;4-(6-((3-(4,4-difluoropiperidin-1-yl)azetidin-1-yl)methyl)-2-(5-fluoro-1H-indol-4-yl)pyrido[3,2-d]pyrimidin-4-yl)morpholine;4-(1-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholine;4-(1-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)piperazin-2-one;2-(1-((2-(1H-indol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;4-(2-(5-fluoro-1H-indol-4-yl)-6-(3-(1,1-dioxo)thiomorpholinoazetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine;4-(2-(5-fluoro-1H-indol-4-yl)-6-(4-(oxetan-3-yl)piperidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine;4-(1-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholine;2-(1-((2-(1H-indol-3-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;and4-(1-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)piperazin-2-one.6. The compound of claim 1 selected from2-(1-((2-(2-methyl-1H-indol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;2-(1-((2-(2-ethyl-2H-indazol-3-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;tert-butyl4-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidine-1-carboxylate;2-(1-((2-(1H-indazol-3-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;4-(2-(5-fluoro-1H-indol-4-yl)-6-(piperidin-4-ylmethyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine;1-(((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)(methyl)amino)-2-methylpropan-2-ol;4-(2-(5-fluoro-1H-indol-4-yl)-6-(1-isopropyl-1H-1,2,4-triazol-5-yl)pyrido[3,2-d]pyrimidin-4-yl)morpholine;N-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)tetrahydro-2H-pyran-4-amine;1-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methylamino)-2-methylpropan-2-ol;4-(6-((3,3-dimethylpyrrolidin-1-yl)methyl)-2-(5-fluoro-1H-indol-4-yl)pyrido[3,2-d]pyrimidin-4-yl)morpholine;N-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)pivalamide;4-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)morpholine;N-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)isobutyramide;1-(4-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)ethanone;1-(4-((2-(5-fluoro-1H-indol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)-2-methylpropan-1-one;(S)-4-(1-((2-(2-(1-methoxyethyl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholine;2-(1-((2-(2-(dimethylamino)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;4-(2-(5-fluoro-1H-indol-4-yl)-6-((tetrahydro-2H-pyran-4-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine;2-(1-((4-morpholino-2-(2-(trifluoromethyl)-1H-benzo[d]imidazol-1-yl)pyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;2-(1-((2-(2-(difluoromethyl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;2-(1-(6-((4-(2-hydroxypropan-2-yl)piperidin-1-yl)methyl)-4-morpholinopyrido[3,2-d]pyrimidin-2-yl)-1H-benzo[d]imidazol-2-yl)acetonitrile;2-(1-((2-(imidazo[1,2-a]pyridin-5-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;N,N-dimethyl-1-(4-morpholino-6-((3-(1,1-dioxo)thiomorpholinoazetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-2-yl)-1H-benzo[d]imidazol-2-amine;(S)-1-(4-((2-(2-(1-methoxyethyl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)ethanone;1-(4-((2-(2-(dimethylamino)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)ethanone;2-(1-((2-([1,2,4]triazolo[1,5-a]pyridin-5-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;N,N-dimethyl-1-(4-morpholino-6-((3-morpholinoazetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-2-yl)-1H-benzo[d]imidazol-2-amine;2-(1-((2-(2-(2-hydroxyethyl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;4-(2-(2-(difluoromethyl)-1H-benzo[d]imidazol-1-yl)-64(3-(1,1-dioxo)thiomorpholinoazetidin-1-yl)methyl)pyrido[3,2-d]pyrimidin-4-yl)morpholine;2-(1-((2-(2-methoxy-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;1-(4-((2-(2-(difluoromethyl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)ethanone;4-(1-((2-(2-(difluoromethyl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholine;2-(1-((4-morpholino-2-(2-(2,2,2-trifluoroethyl)-1H-benzo[d]imidazol-1-yl)pyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;2-(1-((2-(2-(azetidin-1-yl)-1H-benzo[d]imidazol-1-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;2-(1-((4-morpholino-2-(2-(pyrrolidin-1-yl)-1H-benzo[d]imidazol-1-yl)pyrido[3,2-d]pyrimidin-6-yl)methyl)piperidin-4-yl)propan-2-ol;4-(1-((2-(1H-benzo[d]imidazol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholine;and4-(1-((2-(2-methyl-1H-benzo[d]imidazol-4-yl)-4-morpholinopyrido[3,2-d]pyrimidin-6-yl)methyl)azetidin-3-yl)morpholine.7. A pharmaceutical composition comprised of a compound of claim 1 and apharmaceutically acceptable carrier, glidant, diluent, or excipient. 8.The pharmaceutical composition according to claim 17, further comprisinga chemotherapeutic agent.
 9. A kit for treating a condition mediated bythe p110 delta isoform of PI3 kinase comprising a first pharmaceuticalcomposition of claim 7; and instructions for use.